Multiplex genotyping based on the melting temperature of a single locked nucleic acid probe

Ahn, JJ; Kim, JH; Kim, Y; Hong, JY; Kim, GW; Hwang, SY

HERO ID

3539410

Reference Type

Journal Article

Year

2015

Language

English

PMID

26384644

HERO ID 3539410
In Press No
Year 2015
Title Multiplex genotyping based on the melting temperature of a single locked nucleic acid probe
Authors Ahn, JJ; Kim, JH; Kim, Y; Hong, JY; Kim, GW; Hwang, SY
Journal Analytical Biochemistry
Volume 491
Page Numbers 72-74
Abstract Locked nucleic acid (LNA) is a modified RNA nucleotide that can be incorporated at specific positions to generate probes with the desired length, melting temperature (TM), and specificity. Here, we describe a method of multiplex genotyping based on dramatic shifts in the TM of a single dual-labeled LNA probe. Using this method, two varieties of the hairtail fish Trichiurus lepturus can be distinguished from each other, as well as from Trichiurus japonicus, based on a 1- to 2-bp difference in a fragment of mitochondrial cytochrome oxidase subunit 1. The shift in TM was 15 °C for a 1-bp mismatch and 27 °C for a 2-bp mismatch, indicating remarkable specificity. We anticipate that the method will be widely useful in applications such as species identification that require accurate, multiplex, and efficient detection of DNA polymorphisms.
Doi 10.1016/j.ab.2015.09.007
Pmid 26384644
Wosid WOS:000364265200013
Is Certified Translation No
Dupe Override No
Is Public Yes
Language Text English