characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40487_Track-80233: 103166; L40487_Track-79620: 250855

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


status: PUBLIC

tag: attribute

characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

tag: Namespace:The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40509_Track-80050: 258674; L40509_Track-79488: 517882

characteristic: number of reads

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

text: The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40531_Track-80039: 235842; L40531_Track-79574: 472409

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40448_Track-79954: 115669; L40448_Track-79507: 230330

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40463_Track-80207: 93738; L40463_Track-79583: 226937

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40468_Track-80227: 112993; L40468_Track-79621: 272694

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40475_Track-80098: 225096; L40475_Track-79324: 455811

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40444_Track-80057: 173896; L40444_Track-79568: 346925

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40501_Track-79913: 190343; L40501_Track-79451: 380627

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40450_Track-79933: 195758; L40450_Track-79538: 406146

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40484_Track-79658: 941985

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40502_Track-80062: 335789; L40502_Track-79365: 673225

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40458_Track-79659: 820734

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40507_Track-80020: 228844; L40507_Track-79483: 459822

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40522_Track-80165: 132358; L40522_Track-79394: 257105

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40457_Track-80239: 331345; L40457_Track-79632: 809846

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40455_Track-80124: 312486; L40455_Track-79308: 627129

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40481_Track-80052: 224423; L40481_Track-79559: 447516

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40479_Track-80114: 248953; L40479_Track-79450: 496538

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

text: The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40485_Track-80109: 183953; L40485_Track-79500: 369523

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40523_Track-79912: 155144; L40523_Track-79460: 311701

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40489_Track-80010: 152962; L40489_Track-79491: 304677

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40461_Track-79676: 1072746

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40528_Track-79910: 204654; L40528_Track-79432: 409884

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40441_Track-80273: 171190; L40441_Track-79614: 413117

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40465_Track-79960: 151058; L40465_Track-79375: 302581

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40471_Track-80110: 146817; L40471_Track-79302: 293400

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40498_Track-79915: 173615; L40498_Track-79429: 346324

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40512_Track-79945: 132047; L40512_Track-79417: 265826

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40515_Track-80184: 122028; L40515_Track-79435: 245812

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40496_Track-80167: 231610; L40496_Track-79455: 463374

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40500_Track-80131: 179875; L40500_Track-79469: 357407

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40508_Track-80177: 203186; L40508_Track-79370: 405384

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40499_Track-80275: 89066; L40499_Track-79639: 217391

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

text: embryo

text: normal

text: L40456_Track-79968: 374898; L40456_Track-79353: 753646

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP negative

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

text: The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40467_Track-80107: 156000; L40467_Track-79292: 314944

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40647_Track-79652: 221523

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40543_Track-80014: 150688; L40543_Track-79465: 301143

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40562_Track-79998: 163423; L40562_Track-79298: 325247

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40547_Track-80134: 261096; L40547_Track-79307: 525025

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40684_Track-80003: 180120; L40684_Track-79357: 390214

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40673_Track-80089: 228121; L40673_Track-79349: 450073

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40438_Track-80255: 329974; L40438_Track-79594: 596202

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP negative

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40594_Track-80075: 225309; L40594_Track-79314: 445602

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40830_Track-79662: 367122

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 0

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40555_Track-80056: 352218; L40555_Track-79480: 704350

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40642_Track-80095: 103024; L40642_Track-79414: 188633

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40705_Track-79958: 190836; L40705_Track-79458: 384123

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40583_Track-79650: 514109

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40602_Track-80093: 214465; L40602_Track-79283: 426616

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40617_Track-80113: 179822; L40617_Track-79466: 361518

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40685_Track-79977: 143628; L40685_Track-79356: 288229

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

text: embryo

text: normal

text: L40697_Track-80041: 232665; L40697_Track-79440: 468732

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

text: The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40832_Track-79671: 402514

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 0

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40665_Track-79657: 33802

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40620_Track-79921: 222083; L40620_Track-79464: 445047

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40833_Track-79670: 383298

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 0

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40536_Track-80195: 228564; L40536_Track-79392: 459326

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40614_Track-79941: 31836; L40614_Track-79430: 54586

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40597_Track-80172: 176292; L40597_Track-79390: 357003

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40702_Track-80237: 133907; L40702_Track-79579: 296944

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40582_Track-80274: 79741; L40582_Track-79598: 191959

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40600_Track-80061: 239533; L40600_Track-79495: 480507

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40590_Track-79992: 183990; L40590_Track-79382: 365847

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40718_Track-80145: 209445; L40718_Track-79396: 414146

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40706_Track-80147: 176129; L40706_Track-79528: 349898

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40648_Track-80137: 210987; L40648_Track-79393: 414987

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40569_Track-80091: 137177; L40569_Track-79358: 271844

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40655_Track-80009: 261749; L40655_Track-79398: 523189

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40601_Track-80035: 283406; L40601_Track-79423: 569649

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

text: embryo

text: normal

text: L40828_Track-79674: 440703

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 0

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40827_Track-79665: 251981

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 0

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40534_Track-79653: 334103

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40668_Track-80178: 162043; L40668_Track-79433: 318830

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

text: The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40580_Track-80106: 274571; L40580_Track-79536: 551546

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40554_Track-80060: 297984; L40554_Track-79498: 594616

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40616_Track-79997: 218804; L40616_Track-79459: 437274

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40682_Track-80154: 173675; L40682_Track-79383: 346186

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40656_Track-80263: 95474; L40656_Track-79631: 250201

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40577_Track-80252: 208542; L40577_Track-79611: 507253

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40545_Track-79975: 200458; L40545_Track-79567: 401705

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40608_Track-80188: 211345; L40608_Track-79476: 421409

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40686_Track-80234: 74621; L40686_Track-79638: 177968

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40716_Track-80129: 192654; L40716_Track-79535: 383696

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40721_Track-80191: 199821; L40721_Track-79361: 403847

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40533_Track-80055: 148444; L40533_Track-79427: 297521

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40546_Track-79939: 196168; L40546_Track-79384: 394107

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40630_Track-80198: 68987; L40630_Track-79543: 132411

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

text: embryo

text: normal

text: L40831_Track-79660: 407521

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 0

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40558_Track-80122: 113544; L40558_Track-79547: 227788

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

text: The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40671_Track-80132: 220262; L40671_Track-79439: 439341

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40568_Track-80048: 225521; L40568_Track-79486: 451049

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40687_Track-80072: 121787; L40687_Track-79489: 244897

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40695_Track-80045: 265313; L40695_Track-79497: 533553

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40637_Track-80163: 167202; L40637_Track-79475: 330190

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40690_Track-79989: 230613; L40690_Track-79348: 457283

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40700_Track-79955: 348885; L40700_Track-79467: 691275

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40829_Track-79668: 378759

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 0

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40576_Track-79978: 229385; L40576_Track-79290: 462418

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40649_Track-80043: 287079; L40649_Track-79285: 571689

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40658_Track-80040: 132472; L40658_Track-79534: 255855

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40587_Track-80206: 105730; L40587_Track-79514: 206345

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40826_Track-79672: 494410

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 0

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40566_Track-80097: 158994; L40566_Track-79289: 319746

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40639_Track-79654: 4465

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40704_Track-80024: 88209; L40704_Track-79372: 166428

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40699_Track-79982: 164404; L40699_Track-79533: 330918

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

characteristic: description

text: embryo

text: normal

text: L40553_Track-80051: 373406; L40553_Track-79516: 745448

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

text: The Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG)

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40713_Track-79986: 164008; L40713_Track-79436: 329361

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40627_Track-80120: 164900; L40627_Track-79347: 334045

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40674_Track-80066: 186139; L40674_Track-79555: 366423

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40613_Track-80081: 146040; L40613_Track-79288: 293724

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40591_Track-80046: 242361; L40591_Track-79434: 490049

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40605_Track-80228: 263224; L40605_Track-79635: 637829

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40715_Track-79952: 180808; L40715_Track-79570: 363142

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40571_Track-80085: 233288; L40571_Track-79363: 469297

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

characteristic: description

text: embryo

text: normal

text: L40633_Track-80216: 162714; L40633_Track-79590: 377916

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40551_Track-80190: 241009; L40551_Track-79472: 487108

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

text: normal

text: L40680_Track-80130: 172575; L40680_Track-79556: 343716

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

characteristic: description

text: embryo

text: normal

text: L40672_Track-80217: 145162; L40672_Track-79577: 345998

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: embryo

characteristic: developmental stage

text: normal

characteristic: disease

characteristic: genotype

characteristic: gfp mean

text: L40535_Track-79943: 199778; L40535_Track-79542: 392378

characteristic: number of reads

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1


characteristic: External Id

characteristic: INSDC center name

characteristic: INSDC last update

text: public

characteristic: SRA accession

characteristic: Submitter Id

text: ArrayExpress

characteristic: broker name

text: pancreatic cell

text: human

text: embryo

text: normal

text: L40720_Track-80236: 196638; L40720_Track-79589: 472996

text: Homo sapiens

characteristic: organism

text: Homo sapiens

text: inner cell mass

characteristic: organism part

text: GFP low

characteristic: phenotype

text: plate 1

characteristic: plate id

text: human embryonic stem cell

text: Homo sapiens

text: OK

text: Stage 4 Day 1

characteristic: title


characteristic: ENA-CHECKLIST

characteristic: ENA-FIRST-PUBLIC

characteristic: ENA-LAST-UPDATE

characteristic: INSDC first public

text: public

characteristic: INSDC status

text: ArrayExpress

text: pancreatic cell

text: human

characteristic: common name

text: Protocols: hESC-derived pancreatic cells at Stage 4 Day 1 of the pancreatic differentiation protocol (Rezania et al., 2014; Petersen et al., 2017) were dissociated into single cells with TrypLE and resuspended into a FACS buffer containing 0.5% BSA and 2 mM EDTA (Thermo Fisher) in 1X PBS. The cells were index-sorted according to their GFP expression into 384-well plates on a BD FACSAria and snap-frozen. Cells are processed with SMARTseq-2 protocol miniaturized and modified based on Picelli et al. 2013 (doi: 10.1038/nmeth.2639). In short, cells are sorted in 0.5 µl 0.2% Triton X-100 solution with RNase Inhibitor into a 384-well plate, frozen at -80°C. After thawing, the cell lysate is mixed with 0.5 ul of a primer mix (containing 5 mM dNTP (Invitrogen), 0.5 uM dT-primer*, 1 U RNase Inhibitor (NEB)). RNA is denatured (3 minutes 72 °C), followed by reverse transcription (42 °C 90 min + 70°C 15 min) after adding 1.5 ul RT buffer mix (1x superscript II buffer (Invitrogen), 1 M betaine, 5 mM DTT, 6 mM MgCl2, 1 uM TSO-primer*, 2.3 U RNase Inhibitor and 25 U Superscript II). The cDNA is amplified using 1x Kapa HiFi HotStart Readymix (Roche) and 0.1 uM UP-primer under following cycling conditions: 98°C for 3 min, 23 cycles [98°C 20 sec, 67°C 15 sec, 72°C 6 min] and final elongation at 72°C for 5 min. The amplified cDNA is purified using 1x volume of Sera-Mag SpeedBeads (GE Healthcare) and eluted in 12 ul nuclease free water. For library preparation 1.5 µl cDNA is desiccated and rehydrated in 1 ul Tagmentation mix (1x TTBL buffer, 0.1 ul Tagment DNA Enzyme; from TruePrep DNA Library Prep Kit V2 for Illumina; Vazyme) and tagmented (55°C, 5 min), followed by an index PCR with 1x concentrated KAPA Hifi HotStart Ready Mix and 0.7 µM dual indexing primers (72°C 3 min, 98°C 30 sec, 13 cycles [98°C 10 sec, 63°C 20 sec, 72°C 1 min], 72°C 5 min). After PCR, libraries are quantified with AccuBlue Broad range chemistry (Biotium), equimolarly pooled and purified twice with 0.9x volume Sera-Mag SpeedBeads.

characteristic: description

text: embryo

text: normal

text: L40612_Track-80078: 170604; L40612_Track-79421: 338117

text: Homo sapiens

text: Homo sapiens

text: inner cell mass

text: GFP low

text: plate 1

text: human embryonic stem cell

characteristic: progenitor cell type

text: Homo sapiens

text: OK

characteristic: single cell well quality

text: Stage 4 Day 1

characteristic: time

characteristic: well