characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-07-07T00:23:42Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-07-07T00:23:42Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112769168
characteristics.characteristic: External Id
characteristics.text: Istituto di Ricerche Farmacologiche \"Mario Negri\" IRCCS
characteristics.characteristic: INSDC center name
characteristics.text: 2023-07-07T00:23:42Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-07-07T00:23:42Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14769974
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12746:298_R1
characteristics.characteristic: Submitter Id
characteristics.text: 20
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: KPC
characteristics.characteristic: cell_line
characteristics.text: Epcam+; CD45-PDAC cells
characteristics.characteristic: cell_type
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: PDAC tumors were isolated, minced and processed for single cell suspension. Tissues were digested in 2 ml of Digestion Buffer composed by Collagenase IV 3% in RPMI 10% FBS + 1% P/S, DNAasi I 30U/ul, Hepes 10 mM. The cell suspension was incubated for 30 min at 37oC on a rocker. Then, the digestion was stopped adding 1 mL of RPMI 10% FBS + 1% P/S. The cells suspension was filtered through a 100 µm cell strainer and kept on ice for 4 minutes. SMARTer Stranded Total RNA-Seq Kit - Pico Input Mammalian (Clontech Laboratories, Inc., A Takara Bio Company,California, USA) protocol total RNA samples (0.25–10 ng) were reverse-transcribed using random priming into double-stranded cDNA in the presence of a template switch oligo (TSO). When the reverse transcriptase reaches the 5’ end of the RNA fragment, the enzyme’s terminal transferase activity adds non-templated nucleotides to the 3’ end of the cDNA. The TSO pairs with the added non-templated nucleotide, enabling the reverse transcriptase to continue replicating to the end of the oligonucleotide. This results in a cDNA fragment that contains sequences derived from the random priming oligo and the TSO. PCR amplification using primers binding to these sequences can now be performed. The PCR adds full-length Illumina adapters, including dual barcodes for multiplexing. Ribosomal cDNA is cleaved by ZapR in the presence of the mammalian-specific R-Probes. Remaining fragments are enriched with a second round of PCR amplification using primers designed to match Illumina adapters.
characteristics.characteristic: description
characteristics.text: pancreatic cancer
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: in vivo until neoplasm formation
characteristics.characteristic: growth condition
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: pancreas
characteristics.characteristic: organism part
characteristics.text: neoplasm
characteristics.characteristic: sampling site
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: C57BL/6
characteristics.characteristic: strain
characteristics.text: 298_R1
characteristics.characteristic: title