fulltext.study @t Gmail

SNP genotyping through the melting analysis of unlabelled oligonucleotide applied on dilute PCR amplicon

Paper ID Volume ID Publish Year Pages File Format Full-Text
24033 43490 2011 5 PDF Available
Title
SNP genotyping through the melting analysis of unlabelled oligonucleotide applied on dilute PCR amplicon
Abstract

Adaptation of DNA melting analysis for polymorphic single nucleotides (SNPs) genotyping using an unlabeled oligonucleotide probe for polymorphic DNAs under the presence of fluorescent DNA binding dye necessitates a reaction condition where the probe efficiently associates with a target strand that is PCR amplified. We present experimental evidence that application of an unlabeled probe to a dilute PCR amplicon provides a condition such that the fluorescent signals gained subsequently by probe melting are sufficient to discriminate allelic identities. This approach is best exploited by adapting the multiplexing PCR technique in order to cover multiple SNPs for given samples. 3′-end modification of the probe is unnecessary as the amplicon dilution step provides a way of inactivating the polymerase through divalent cation chelation. With the use of low-cost reagents and ordinary laboratory equipment, this method offers a rapid, simple and cost-efficient way of SNP genotyping.

Keywords
SNP, single nucleotide polymorphism; GWAS, genome-wide association study; UOP, unlabelled oligonucleotide probe; bps, base pairs; TM, melting temperatureSNP; Genotyping; DNA melting
First Page Preview
SNP genotyping through the melting analysis of unlabelled oligonucleotide applied on dilute PCR amplicon
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 154, Issue 4, 20 July 2011, Pages 321–325
Authors
, , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us