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After sample quality control, genomic DNA is fragmented to the desired size using g-TUBEs (Covaris, USA) according to the library fragment size. The target DNA fragments are enriched and purified with AMPure PB magnetic beads, and fragmented DNA is quality checked using 0.7% agarose gel electrophoresis. Library construction is performed on qualified DNA samples using the SMRTbell™ Express Template Prep Kit 2.0. Exonuclease is used to remove single-stranded DNA ends, followed by damage repair, end repair, and Poly-A tail addition to the fragmented DNA, and then the hairpin adapters are ligated. The SMRTbell library undergoes nuclease digestion and purification using the SMRTbell Enzyme Cleanup Kit. Target fragments are then selected using the BluePippin device (Sage Science, USA) and purified to complete the library. Finally, an Agilent 2100 Bioanalyzer (Agilent Technologies, USA) is used to check the library fragment size. |
WGS |
GENOMIC |
PolyA |
SINGLE
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