title: GSM8597001: Control-3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA is isolated using TRIzol Universal and chloroform. Afetr centrifugation, the aqueous phase containing RNA is collected and precipitated using isopropanol. After precipitation and centrifugation, RNA forms a pellet, which is washed to remove impurities and dissolved in DEPC water. RNA-seq library preparation follows the protocol of the Illumina TruSeq™ RNA Sample Prep Kit. Poly-A mRNA is enriched using Oligo(dT) magnetic beads, then fragmented ultrasonically. Using these mRNA fragments, first-strand cDNA is synthesized with random primers and M-MuLV reverse transcriptase, followed by RNA degradation with RNase H. Second-strand cDNA synthesis is done with DNA Polymerase I. The double-stranded cDNA is purified, end-repaired, A-tailed, and sequencing adapters are added. AMPure XP beads select ~200 bp fragments, followed by PCR amplification and another purification to yield the final library.