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  • Optimization of the cDNA-PCR sequencing protocol​

    I'm using the cDNA-PCR Sequencing Kit (SQK-PCS111) to sequence full-length transcripts from human brain tissue. I have followed the protocol from Oxford Nanopore Technologies and I have obtained about 10 ng of cDNA per sample. However, when I run the QC step using the Qubit and the TapeStation, I see a very low concentration of cDNA (around 1 ng/ul) and a very broad size distribution (from 200 bp to over 10 kb). How can I optimize the cDNA-PCR sequencing protocol to get higher yield and quality of cDNA?

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