| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
CCS-hifi sequencing：All sequencing libraries were prepared using SMRTbell Template Prep Kit v.1.0 (Pacific Biosciences Ref. No. 100-259-100). To minimize ligation chimeras, hairpin adapters were ligated overnight at 500× molar excess to the genomic fragment molecules. Sequencing primers were conditioned by heating to 80 °C for 2 min and rapidly cooled to 4 °C. The sequencing primer was annealed to the template at a molar ratio of 20:1 (primer:template) for 30 min at 20 °C. After primer annealing, polymerase was bound to the primed template at a molar ratio of 10:1 (polymerase:template) for 4 h at 30 °C. Polymerase-bound samples were then kept at 4 °C before use. Before sequencing, excess unbound polymerase was removed by incubating the complexes for 5 min with 0.6× (vol:vol) AMPure PB beads (Pacific Biosciences Ref. No. 100-265-900) at room temperature. Beads were not washed with 80% ethanol. After removing the free polymerase in the supernatant, the polymerase-bound complexes were eluted with MagBead Binding Buffer v.2 (Pacific Biosciences Ref. No. 101-046-400). |
WGS |
GENOMIC |
other |
SINGLE
|
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