GSM5343241: mouse skin stromal cells after injury; Mus musculus; RNA-Seq
All samples were purified with SPRI beads (ratio 1: 0.8) after pre-amplification. Final elution was in 10 µL of nuclease-free water (Invitrogen). Later cDNA was quantified using the Quant-iT PicoGreen double stranded DNA Assay Kit (Thermo Fisher). A High Sensitivity DNA chip (Agilent Bioanalyzer), was used to verify size distributions. Samples that passed the quality and quantity control parameter, were used for constructing Nextera XT libraries from 0.8 ng of pre-amplified cDNA. Later, 3 'end sequences were enriched with a custom made P5 primer (P5NEXTPT5, IDT), during PCR of the libraries. Following this, Libraries were pooled and selected based on their size using 2% E-Gel Agarose EX Gels (Life Technologies); a size range from 300bp to 800 bp was cut out and extracted using the MinElute kit (Qiagen). All Procedures were in accordance with the manufacturer's recommendations. An Illumina HiSeq 1500 instrument was used to sequence libraries. Paired-end sequencing of libraries was performed on high output flow cells. To generate molecular and cellular barcodes, 16 bases were sequenced with the 1st read, 50 bps sequenced in the 2nd read into the cDNA fragment, and 8 bases were read to obtain the i7 barcode.
GEO
GSM5343241