GSM2544624: HT1080_Low Density_10cells/mm^3; Homo sapiens; RNA-Seq
The matrices were exposed to cell lysis buffer (Trizol) and mechanically broken down using a syringe. Total RNA isolation was performed with RNA MiniPrep kit (Zymo Research). mRNA was selected from total RNA using phosphorylated oligo-dT, and single-strand cDNA was generated from mRNA with SuperScript Reverse Transcriptase III. Double-strandcDNA generation was performed. The End-It DNA End-Repair kit (which includes End-it buffer, 1mM dNTPs, 1mM ATP, total enzyme mix) and T4 DNA ligase were combined for the DNA end-blunting, 5'-end phosphorylation, and ligation. phi29 DNA polymerase-based whole DNA pool amplication was adopted for mRNA transcriptome amplifcation. cDNA libraries were prepared for sequencing using the standard protocol suggested by Illumina Nextera XT DNA Library Prep Kit