GSM5462729: PG7_44: 3211 normal; Homo sapiens; RNA-Seq
Total RNA was extracted by using the RNAiso Plus (Takara Bio, Japan). Extracted RNA was purified by using the RNeasy Mini Kit accompanied with the DNase I (Qiagen) treatment. mRNA was enriched by using poly-T oligo-attached magnetic beads, followed by mRNA fragmentation by acoustic shearing. First-strand cDNA was synthesized by using reverse transcriptase and random hexamers, and second-strand cDNA by using DNA polymerase I and RNase H. cDNA was subject to adapter ligation, and then enriched with PCR to prepare cDNA library. cDNA libraries were sequenced on HiSeq 2000 (Illumina) to obtain around 100 million paired-end reads (2 x 101 bp).