RNA-seq of TSCC cells isolation from mice xenograft treated with H2O
After the sample test is qualified, the library is constructed. The main process is as follows:(1) enrichment of eukaryotic mRNA by magnetic beads with Oligo (dT).(2) join Fragmentation Buffer to interrupt mRNA randomly.(3) using mRNA as template, the first cDNA strand was synthesized with six-base random primers (random hexamers), and then buffered.The second cDNA chain was synthesized by liquid, dNTPs, RNase H and DNA polymerase I, and cDNA was purified by AMPure XP beads.(4) the purified double stranded cDNA was repaired by terminal repair, A tail was added and sequenced splice was connected, and then AMPure XP beads was used for slicing.Segment size selection.(5) finally, the cDNA library was obtained by PCR enrichment.