characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-10-14T05:00:41Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-10-14T05:00:41Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA113991934
characteristics.characteristic: External Id
characteristics.text: Department of Systems Biology School of Life Sciences Southern University of Science And Technology
characteristics.characteristic: INSDC center name
characteristics.text: 2023-10-14T05:00:41Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-10-14T05:00:41Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS15975783
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-13115:zf_3.3hpf_plate1_369
characteristics.characteristic: Submitter Id
characteristics.text: 3.3
characteristics.unit: hour
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: not collected
characteristics.characteristic: collection date
characteristics.text: zebrafish
characteristics.characteristic: common name
characteristics.text: Protocols: Staged embryos were transferred into 10cm dish on ice. The chorion was physically disrupted and removed by tweezers. Then the majority of the yolk was physically removed using a tweezer. The remaining embryonic cells were washed with PBS and incubated in the deyolk buffer. When single cells became dissociated, they were transferred into a new eppendorf tube and washed with PBS for three times. Nuclei acid was extracted first using a Qiagen PCR purification column. Then 1× VAHTS DNA cleaning beads were used to remove primer leftover and eluted in 20 μl Nuclease-free Water. 50,000 single cells were resuspended in 50 uL RSB + 0.01% digitonin 0.1% Tween-20 and 0.1% NP-40 and incubated on ice for 3 minutes. Then 1 mL of cold RSB + 0.1% Tween-20 (without digitonin and NP-40) was added. The permeabilised cells were centrifuged at 300 g 4 C. After removing the supernatant, 50 ul tagmentation mix was used to resuspend the cell pellet and incubated at 37C on a thermomixer for 30 minutes. Then 50 ul 20mM EDTA was added to stop the reaction. Single cells were sorted into each well of a plate containing distinct well index primers using FACS. 2x NEBNext PCR Master Mix was added into each well to perform PCR for the library construction. The PCR condition is: 72 C 5 minutes, 98 C 2 minutes, then 12 cycles of [98 C 10 seconds, 63 C 30 seconds, 72 C 20 seconds].
characteristics.characteristic: description
characteristics.text: blastula high
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: not collected
characteristics.characteristic: geographic location (country and/or sea)
characteristics.text: 3.3
characteristics.unit: hour
characteristics.characteristic: hours post fertilisation (hpf)
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Danio rerio
characteristics.characteristic: organism
characteristics.text: Danio rerio
characteristics.characteristic: organism
characteristics.text: embryonic structure
characteristics.characteristic: organism part
characteristics.text: Danio rerio
characteristics.characteristic: scientific_name
characteristics.text: mixed
characteristics.characteristic: sex
characteristics.text: Tubingen
characteristics.characteristic: strain
characteristics.text: zf_3.3hpf_plate1_369
characteristics.characteristic: title