Amplicon sequencing analysis of on-target sites in trβ crispants

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Fig. 2. Amplicon sequencing analysis of on-target sites in trβ crispants. Amplicon sequencing analysis of on-target sites in trβ crispants. (A) Image of trβ crispants at stage 66. Genomic DNA was extracted from whole bodies of eight trβ crispants and the sgRNA target region was PCR-amplified using barcoded primers. PCR amplicons were subjected to amplicon sequencing according to Materials and Methods. Numbers indicate each crispant that was analyzed by amplicon sequencing in B. (B) The results of wild-type and mutant allele reads in each trβ crispant. All sequenced reads were classified into three groups; wild-type reads, frameshift and/or splice donor site mutation reads, in-frame mutation reads. Yuto Sakane et al. Biology Open 2018;7:bio030338 © 2018. Published by The Company of Biologists Ltd