Rat1
An appropriate amount of interrupting reagent was added to the obtained mRNA, which was allowed to break at a high temperature. Single-stranded cDNA was synthesized using the interrupted mRNA as a template. A double-strand synthesis reaction system was then configured to synthesize double-stranded cDNA. Kits were used to purify and recover cDNA, repair sticky ends, add base " A " to the 3 'end of cDNA and ligate linkers, followed by fragment size selection and amplification by PCR. The constructed libraries were examined by Agilent 2100 Bioanalyzer and ABI StepOne Plus Real-Time PCR system and qualified for sequencing.