Cas12a3_NT1h
Extract total RNA from the sample, then remove rRNA, fragment mRNA, and use random primers to reverse transcribe to synthesize one strand of cDNA. During the synthesis of the second strand, dNTPs reagent is replaced with dUTP, so that the bases in the second strand of cDNA include A/U/C/G. The double-stranded cDNA structure has a sticky end, which is ligated to a Y-shaped connector. Use UNG enzyme to digest the second strand of cDNA, thereby making the library contain only the first strand of cDNA.