RNA-Seq of Macrobrachium nipponense
After enriching eukaryotic mRNA with polyA tail using magnetic beads with Oligo(dT), the mRNA is fragmented with buffer. Using fragmented mRNA as a template and random oligonucleotides as primers, the first strand of cDNA was synthesized in the M-MuLV reverse transcriptase system, and then RNaseH was used to degrade the RNA chain, and dNTPs were used as the primer in the DNA polymerase I system. The raw materials are used to synthesize the second strand of cDNA. The purified double-stranded cDNA is end-repaired, A-tailed, and connected to sequencing adapters. AMPure XP beads are used to screen about 200 bp of cDNA, PCR amplification is performed, and AMPure XP beads are used again to purify the PCR product to finally obtain a library.