An improved heteroduplex analysis for rapid genotyping of SNPs and single base pair indels

被引：4

作者：

Fan, Jiangbo ^{[1
,2
]}

Xia, Ye ^{[1
]}

Wang, Guo-Liang ^{[1
,2
]}

机构：

[1] Ohio State Univ, Dept Plant Pathol, Columbus, OH 43210 USA

[2] Chinese Acad Agr Sci, Inst Plant Protect, State Key Lab Biol Plant Dis & Insect Pests, Beijing 100193, Peoples R China

来源：

BIOTECHNIQUES | 2019年 / 67卷 / 01期

关键词：

CRISPR; genotyping; heteroduplex assay; iHDA; improved heteroduplex assay; single base pair insertion/deletion; single nucleotide polymorphism; GENOME; MUTAGENESIS; MUTATIONS; MAP;

D O I：

10.2144/btn-2019-0012

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

SNPs and single base pair (SBP) insertion/deletions (indels) are not only the most abundant genetic markers for genetic mapping and breeding selection, but also always occur in the mutants generated from chemical mutagenesis or CRISPR/ Cas9-mediated genome editing. Most of the current SNP and SBP indel genotyping methods are time-consuming and/or require special equipment or reagents. Here, we describe an improved heteroduplex analysis method, named iHDA, that can readily discriminate SNP and SBP indel alleles with specially designed DNA probes that harbor a couple of nucleotides adjacent to the SNP site. By hybridizing with the same probe, SNP and SBP indel alleles form different heteroduplexes, differing in bulge size, which show different mobility on a polyacrylamide gel. Therefore, iHDA is an easy, fast and inexpensive method for SNP and SBP indel genotyping.

引用

页码：6 / 10

页数：5

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