Screening of novel functional genes from Bacillus thuringiensis
The DNA fragments longer than or equal to 10 Kb were retrieved by BluePippin DNA fragment screening system (Sage Science, USA) according to the instructions. Then, the steps of SQK-LSK108 1D ligation Genomic DNA library preparation as follow:   First, DNA damage was repaired by NEBNext FFPE DNA Repair Mix (New England Biolabs Ltd., USA). After purification, the NEBNext End Repair/ DA-tailing Module Kit (New England Biolabs Ltd., USA) was used to Repair the ends of the DNA fragments. Then, Neb Blunt/Ta Ligase Master Mix Kit (New England Biolabs Ltd., USA) was used for joint connection, and quantification with Qubit was performed again for quantitative detection of the DNA library. The constructed library was loaded into the Flow Cell on a Nanopore GridION X5 device for real-time single-molecule sequencing.
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