Premium
Generation of co‐dominant PCR‐based markers by duplex analysis on high resolution gels
Author(s) -
Hauser MarieTheres,
Adhami Farzaneh,
Dorner Maria,
Fuchs Esther,
Glössl Josef
Publication year - 1998
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1046/j.1365-313x.1998.00271.x
Subject(s) - heteroduplex , biology , genetics , single nucleotide polymorphism , cleaved amplified polymorphic sequence , microbiology and biotechnology , ecotype , high resolution melt , gene , polymerase chain reaction , computational biology , restriction fragment length polymorphism , genotype
Summary Rapid and efficient procedures for the detection of sequence polymorphisms are essential for chromosomal walking and mutation detection analyses. While DNA chip technology and denaturing high‐performance liquid chromatography (DHPLC) are the methods of choice for large scale facilities, small laboratories are dependent on simple ready‐to‐use techniques. We show that heteroduplex analysis on high resolution gel matrices efficiently detects sequence polymorphism differing as little as a single base pair (e.g. single‐nucleotide polymorphism, SNP) with standard laboratory equipment. Furthermore, the matrices also discerned differences between homoduplexes, a prerequisite for co‐dominant markers. The markers thus generated are referred to as duplex analysis markers. We designed PCR primers for 36Arabidopsis thalianaloci ranging in length from 230 bp to 1000 bp. Among three ecotypes, more than half (n= 19) of the loci examined were polymorphic; five of which contained three different alleles. This simple, high resolution technique can be used to rapidly convert sequence tagged sites into co‐dominant PCR‐based molecular markers for fine‐scale mapping studies and chromosomal walking strategies as well as for the detection of mutations in particular genes.