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cDNA library construction and online sequencing

QW Qichao Wu
FZ Fengqi Zang
XX Xiaoman Xie
YM Yan Ma
YZ Yongqi Zheng
DZ Dekui Zang
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A cDNA library was constructed using qualified samples. The SMARTer™ PCR cDNA Synthesis Kit was used for the synthesis of full-length cDNA from mRNA. The full-length cDNA fragments were screened using BluePippin and then amplified again by PCR. Subsequently, end repair of the full-length cDNA was performed, and the dumbbell-shaped SMRT adapter was connected. After exonuclease digestion, BluePippin was used for secondary screening to obtain a cDNA library. Qubit 2.0 and Agilent 2100 were used to accurately quantify and detect the library size, respectively. After the libraries passed quality control, full-length transcriptome sequencing was performed using PacBio RSII according to the target offline data volume.

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