GSM5737629: KGN cells, NC 3; Homo sapiens; RNA-Seq
The KGN cells were ground into a fine powder before RNA extraction. Total RNA was treated with RQ1 DNase (Promega) to remove DNA. For each sample, 0.5μg of total RNA was used for RNA-seq library preparation using the KAPA Stranded mRNA-Seq Kit for Illumina® Platforms (KK8544). Polyadenylated mRNAs were purified, fragmented and converted into double strand cDNA. Following the end repair and A tailing, the DNA fragments were ligated to the Diluted Roche Adaptor (KK8726). After purifying and size fractioning to 300-500bps, the ligated products were amplified and purified, quantified and stored at -80℃ before sequencing. The strand marked with dUTP (the 2nd cDNA strand) was not amplified, allowing for strand-specific sequencing. The libraries were prepared for high-throughput sequencing following the manufacturer's instructions for the Illumina NovaSeq 6000 system with 150 nt paired-end sequencing.