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Randomized PAM depletion of AsCas12a

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https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE123443
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We report the PAMs of AsCas12a using a cell-free TXTL-based cleavage assay. By adding randomized PAM library and AsCas12a-gRNA in vitro, functional PAM sequences were cleaved, while non-functional PAMs remained. By amplifying the non-cleaved DNA, we use next-generation sequencing to analyze the depletion of functional PAMs of AsCas12a. Uncleaved DNA from the randomized PAM library was amplified using Illumina Nextera Indices
创建时间:
2019-04-22
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