mRNA-seq of human liver sinusoidal cell
Firstly, purifying the poly-A containing mRNA molecules using poly-T oligo-attached magnetic beads. Following purification, the cleaved RNA fragments are copied into first strand cDNA using reverse transcriptase and random primers. This is followed by second strand cDNA synthesis and added a single 'A' base to these cDNA fragments and subsequent ligation of the adapter. The products are then purified and enriched with PCR amplification. Then quantified the PCR yield by Qubit and pooled samples together to make a single strand DNA circle (ssDNA circle), which gave the final library.