GSM4506350: HMDM M1 rep20; Homo sapiens; RNA-Seq
RNA samples were extracted using All Prep DNA/RNA/miRNA Universal Kit (Qiagen, Valencia, CA). Extracted RNA samples underwent quality control (QC) assessment using the Agilent Bioanalyzer (Agilent, Santa Clara, CA) With a minimum of 300 ng input RNA, we generated first-strand cDNA using random hexamer-primed reverse transcription, followed by second-strand cDNA synthesis using RNase H and DNA polymerase, and ligation of sequencing adapters using the TruSeq RNA Sample Preparation Kit (Illumina, San Diego, CA). Modification of protocol include (1) fragmentation time is decreased from 8 to 6 min to ensure libraries were > 100 bp long and (2) PCR amplification is limited to 12 cycles. To minimize lane and batch effects in RNA-Seq experiments, barcoded libraries were mixed and pooled in equimolar (2 nmol/L) amounts and diluted to 4 pmol/L for cluster formation on a single flow cell lane, followed by 100 bp paired-end sequencing on Illumina's HiSeq 2500.
GEO Accession
GSM4506350