Differential gene expression induced by overexpression of ECI2
(1) Enrichment of eukaryotic mRNA with magnetic beads with Oligo (dT); (2) Break the mRNA into short fragments; (3) Synthesize the first strand of cDNA using the fragmented mRNA as a template and a six-base random primer; (4) Add buffer, dNTPs and enzymes to synthesize the second cDNA strand (add dTTP, dATP, dGTP and dCTP for general library construction and dUTP, dATP, dGTP and dCTP for strand-specific library construction); (5) The obtained double-stranded cDNA was purified, end repaired, A-tailed and connected to sequencing junction, followed by fragment size selection; (7) PCR amplification and enrichment of cDNA library (special PCR amplifying enzyme is used for strand-specific library construction and PCR enrichment of U-free template).