Allele-specific Holliday junction formation: a new mechanism of allelic discrimination for SNP scoring.

Abstract:

:We report here a new mechanism for allelic discrimination--allele-specific Holliday Junction formation. The Holliday Junction (HJ) is a unique DNA structure that can be formed in a sequence-nonspecific manner by routine PCR. To cause the PCR-based HJ formation to occur in an allele-specific manner, the PCR primers are manipulated such that an extra mismatch next to a SNP of interest is introduced between a target and a reference amplicon and a GC-clamp is added. Based on this new mechanism, novel SNP genotyping methods were developed, including a homogeneous fluorescence polarization (FP) competition assay that requires neither labeled primers/probes nor expensive enzymes/substrates. Using this novel genotyping technology, we were able to convert >95% of SNP sequences into genotyping assays that work well under a universal set of assay conditions and achieved 100% accuracy in clinical samples.

journal_name

Genome Res

journal_title

Genome research

authors

Yang Q,Lishanski A,Yang W,Hatcher S,Seet H,Gregg JP

doi

10.1101/gr.997703

subject

Has Abstract

pub_date

2003-07-01 00:00:00

pages

1754-64

issue

7

eissn

1088-9051

issn

1549-5469

pii

13/7/1754

journal_volume

13

pub_type

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