In many diploid organisms, the majority mutations induced by clustered regularly interspaced short palindromic repeats (CRISPR)-mediated genome editing are non- chimeric, including biallelic, homozygous, and heterozygous mutations. Direct Sanger sequencing of the PCR amplicons containing non-homozygous mutations superimposes sequencing chromatograms, displaying overlapping peaks beginning from the mutation sites. In this chapter we describe the degenerate sequence decoding (DSD) strategy and its automatic web-based tool, DSDecodeM, for decoding the Sanger sequencing chromatograms from different types of targeted mutations. DSDecodeM, as a convenient and versatile tool, can considerably facilitate the genotyping work of CRISPR-induced mutants.
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This work is supported by grants from Guangdong Province Public Interest Research and Capacity Building Special Fund (2015B020201002) and the Ministry of Agriculture of the People’s Republic of China (2016ZX08010-001, 2016ZX08009-002) and the Postdoctoral Science Foundation of China (2016 M602480).
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