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After the qualified RNA (or polyA+RNA) is detected, the first cDNA strand is synthesized by SMARTer PCR cDNA Synthesis Kit. Once the SMARTScribe Reverse Transcriptase is added to the 5 'end of mRNA, The end repair transferase adds several additional base sequences to the 3 'end of the cDNA, which serve as a binding point for oligonucleotide synthesis, extending the first strand of the cDNA to the end of the oligonucleotide, producing a universal primer sequence that facilitates the synthesis of the second cDNA chain and PCR amplification, thus performing PCR amplification. The principle diagram of double-stranded cDNA synthesis is as follows: |
RNA-Seq |
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