GSM8958685: Lot 6 Nf1-/- DNp53 male transformed astrocytes, replicate 3; Mus musculus; RNA-Seq
Total RNA was isolated from Qiagen's Rneasy Plus Mini Kit (74136). Transcripts are annealed to primers containing a 16-nucleotide sequence unique to each sample (sample barcode), a unique molecular identifier, and a poly A 3' end (Integrated DNA Technologies). First-strand cDNA synthesis of barcoded transcripts was performed using Maxima H Minus (Invitrogen) with a template switch oligo (TSO). Hybrid transcript:cDNA was pooled and exonuclease treated (New England Biotech, M0293), followed by subsequent cDNA amplification (KAPA HiFi HotStart). Library construction was performed with 2 ng of amplified cDNA using the Illumina Nextera XT Protocol.