title: GSM9253275: Renca cells, passage 17, rep2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted using Trizol reagent. RNA purity was assessed with a NanoPhotometer® spectrophotometer (IMPLEN, CA, USA), concentration was measured using a Qubit® 2.0 Fluorometer (Life Technologies, CA, USA), and integrity (RIN value) was evaluated with an Agilent 2100 Bioanalyzer (Agilent Technologies, CA, USA). After quality control, mRNA was enriched using Oligo(dT) magnetic beads, and first-strand cDNA was synthesized with M-MuLV Reverse Transcriptase. RNA templates were degraded by RNase H, and second-strand cDNA was synthesized. Subsequent steps included end repair, poly-A tailing, adapter ligation, and size selection (~200 bp) using AMPure XP beads, followed by PCR amplification for library construction. RNA libraries for RNA-seq were prepared using New England Biolabs (#E7530 ) following manufacturer's protocols.