GSM6592120: gdT, TGFB, resting, 3; Homo sapiens; RNA-Seq
All samples were stored at -80C. Total RNA extraction using RNeasy mini kit (Qiagen) according to the manufacturer's instructions. The quality of RNA was controlled by an Agilent 2100 Bioanalyzer. RNAs were qualified if the 28S/18S ratio was more than 1.5. Libraries were prepared from 10 ng of total RNA. The mRNA poly(A) tail was tagged with universal adapters, well-specific barcodes and unique molecular identifiers (UMIs) during template-switching reverse transcriptase. Barcoded cDNAs from multiple samples were then pooled, amplified and tagmented using a transposon-fragmentation approach that enriches for 3'ends of cDNA. A library of 350-800 bp was run on an Illumina HiSeq 2500 using a TruSeq Rapid SBS Kit (Illumina, San Diego, CA, USA) 3' sequencing RNA 3'-DGE-Seq