GSM1314603: Knockout; Mus musculus; RNA-Seq
Postnatal SPCs were isolated from 8dpp testes. RNA was extracted using Ambion Tri-Reagent and two microgram RNA was used to create the construction of sequencing libraries. Two microgram RNA was purified by using poly-T oligo-attached magnetic beads and mRNA was fragmented into small pieces using divalent cations under elevated temperature. The cleaved RNA fragments were copied into first strand cDNA using reverse transcriptase and random primers, followed by second strand cDNA synthesis using DNA Polymerase I and RNase H. These cDNA fragments were end-repaired, a single A base addition, and adapter ligation. The products were then purified and enriched with PCR to create the final cDNA library. Paired-end 100 nucleotide reads from each mRNA library were obtained using HiScanSQ (Illumina Inc., CIC bioGUNE, Bilbao, Spain).
GEO Accession
GSM1314603