RNA-seq of MDA-MB-231 cells
In this project, mRNA with polyA structure in total RNA was enriched by Oligo(dT) magnetic beads, and the RNA was broken into fragments of about 300bp in length by ion interruption. The segment with a length of 300bp is selected because the length of the joint is fixed. If the length of the interrupted segment is short, the proportion of the joint sequence will be high, thus reducing the proportion of effective data. If the length of the interrupted fragment is long, it is not conducive to the generation of clusters in the process of on-machine sequencing. Using RNA as template, 6-base random primers and reverse transcriptase were used to synthesize the first strand cDNA, and the second strand cDNA was synthesized using the first strand cDNA as template.