GSM5176818: 24h PMA-treated_replicate 1; Homo sapiens; RNA-Seq
Total RNA extraction was carried out on 5x106 cells treated with PMA for 6 and 24 hours using TRIzol according to manufacturer's protocol. Reverse-transcription with oligo-dT primer (unanchored) supplemented with small amounts of 3'-azido-nucleotides (AzVTPs) was done to initiate cDNA synthesis from within the poly(A) tail ensuring stochastic termination upstream of the poly(A) tail. Resulting 3'-azido-blocked cDNA molecules were purified, 'click-ligated' and amplified to generate a cDNA sequencing library. These libraries were pooled and sequenced using the manufacturer's standard operating procedures on a NextSeq 550 obtaining 1×150 bp (130M) single-end reads. Poly(A)-ClickSeq
GEO
GSM5176818