GSM9421135: KYSE150-hypoxia-oxamate-NALA-rep2; Homo sapiens; RNA-Seq
RNA purification, reverse transcription, library construction and sequencing were performed in Mingma Technologies at Shanghai according to the manufacturer's instructions. The agarose gel electrophoresis was used to detect the integrity of the extracted RNA. And NanoDrop was used to measure the concentration of RNA. The mRNAs containing oligoDT were enriched from the total RNA by magnetic beads, and the captured mRNAs were fragmented, then the reverse transcriptase was used to synthesize the strands of cDNA and repair the end of the transcripts product. An “A” base was added to the 3' end of the product and the sequencing adapters were connected to the ends. The incomplete ligation products and empty linker self-ligation products were removed by the ligation products purification, and the primers complementary to the linker sequence were used for PCR amplification. Finally, the sequencing library was purified by magnetic beads.