靶向测序建库-多重PCR法 Fragment DNA- 1.End repair 5 End repair enzyme P1 adapter 5' Barcode lon Xpress Barcode adapter 5'去磷酸 化 po.5 P1Adapter: 2.Ligation of adaptors 5-CCACTACGCCTCCGCTTTCCTCTCTATGGGCAGTCGGTGAT3 3'-T*T*GGTGATGCGGAGGCGAAAGGAGAGATACCCUTCAGCCACIA-5' DNA ligase enzyme Nick lon Xpress"M Barcode adapter: DNA insert 5-CCATCTCATCCCT+G*CGTGTCTCCGACTCAGNNNNNNNNN GA马 3'-C-GCACAGAGGCTGAGTCNNNNNNNNN a*"indicates a phosphorothioate bond,for protectjon 5' from nucleases and to preserve the directionality of adapter Nick repair polymerase.dNTP ligation. Nick GAT=barcode adapter sequence;required to minimize adapter ligation efficiency bias and prevent a two-mer C ncorporation at the end of the barcode sequence. 2024/8/8 30 2024/8/8 30 靶向测序建库-多重PCR法