Enhanced discrimination of single nucleotide polymorphisms using 3' nucleotide differences in ligase detection reaction probes.

Abstract:

:The ligase detection reaction (LDR) is a highly specific genotyping method for single nucleotide variations. Although LDR typically discriminates single nucleotide polymorphism (SNP) alleles at the 3' end of so-called LDR discriminating probes, we designed probes in which the position of nucleotide differences for discrimination was shifted to the second and third nucleotides from the 3' end. Using the 3'-modified probes, we targeted SNPs of the human ABO group and investigated the specificity and efficiency of ligation by a universal LDR assay. We demonstrated that one or two nucleotide shifts of differences in discriminating probes improve the allele balance in detecting both base substitutions and short deletions. In regard to short deletions, moreover, the shifts of nucleotide differences in discriminating probes form the perfect-machted or multiple-mismatched structures (the bulge structures) in the discriminating probe-target DNA duplex and may contribute to enhance ligation efficiency.

journal_name

Mol Cell Probes

authors

Asari M,Omura T,Maseda C,Shiono H,Tasaki Y,Matsubara K,Shimizu K

doi

10.1016/j.mcp.2010.08.005

subject

Has Abstract

pub_date

2010-12-01 00:00:00

pages

381-6

issue

6

eissn

0890-8508

issn

1096-1194

pii

S0890-8508(10)00062-9

journal_volume

24

pub_type

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