RNA-Seq of Heterococcus viridis: collected in day 5 at 5 C replicate 1
Using fragmented mRNA as a template and random oligonucleotides as primers, the first strand of cDNA is synthesized in the M-MuLV reverse transcriptase system. Subsequently, the RNA strand is degraded by RNase H, and the second strand of cDNA is synthesized using dNTPs as substrates in the DNA polymerase I system. Purified double-stranded cDNA undergoes end repair, A-tailing, and sequencing adapter ligation. cDNA fragments of approximately 250-300 bp are selected using AMPure XP beads, amplified via PCR, and purified again with AMPure XP beads to yield the final library.