scholarly journals Allele-specific PCR assay to genotype SNP rs7903146 in TCF7L2 gene for rapid screening of diabetes susceptibility

2008 ◽  
Vol 52 (8) ◽  
pp. 1362-1366 ◽  
Author(s):  
Ludmila Alves Sanches Dutra ◽  
Patrícia Godoy Garcia Costa ◽  
Lara Franciele Ribeiro Velasco ◽  
Angélica Amorim Amato ◽  
Gustavo Barcelos Barra
Keyword(s):  
Rapid Screening ◽  
Complete Agreement ◽  
Diabetes Susceptibility ◽  
Specific Pcr ◽  
Allele Specific ◽  
Reverse Primer ◽  
Specific Amplification ◽  
Common Reverse Primer ◽  
Specific Pcr Assay ◽  
Allele Specific Pcr

OBJECTIVE: The aim of the present study is to validate a rapid and simple allele-specific PCR that genotypes TCF7/L2 rs7903146 (C/T) polymorphism with standard PCR instruments. METHODS: Two forward primers with variations in their 3' nucleotides were designed in such a way that each was specific for one of the two variants. They were combined with a common reverse primer into two PCR reactions. Specific amplification indicates the presence of the allele. One hundred and four DNA samples were genotyped by this method. To evaluate the assay, the polymorphism spanning region of 63 DNA samples representing the three possible genotypes was sequenced. RESULTS: The rs7903146 allele assignments derived from the allele-specific PCR were in complete agreement with sequencing. CONCLUSIONS: The assay described here is a suitable strategy for the TCF7/L2 rs7903146 (C/T) genotyping also allowing rapid and reliable identification.

Development of a multiplex allele-specific PCR assay for rapid screening of Lactoferrin TATA box polymorphisms

2012 ◽  
Vol 22 (6) ◽  
pp. 1241-1244
Author(s):  
Reza Valadan ◽  
Hassan Sharifiyazdi ◽  
Abdolah Mirzaei ◽  
Keshvad Hedayatian
Keyword(s):  
Tata Box ◽  
Rapid Screening ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Allele Specific ◽  
Specific Pcr Assay ◽  
Allele Specific Pcr

Simple and Rapid Detection of Factor V Leiden by Allele-specific PCR Amplification

1996 ◽  
Vol 75 (05) ◽  
pp. 757-759 ◽  
Author(s):  
Rainer Blasczyk ◽  
Markus Ritter ◽  
Christian Thiede ◽  
Jenny Wehling ◽  
Günter Hintz ◽  
...  
Keyword(s):  
Direct Sequencing ◽  
Activated Protein C ◽  
Factor V Leiden ◽  
Pcr Amplification ◽  
Factor V ◽  
Specific Pcr ◽  
Pcr Rflp ◽  
Allele Specific ◽  
Specific Amplification ◽  
Allele Specific Pcr

SummaryResistance to activated protein C is the most common hereditary cause for thrombosis and significantly linked to factor V Leiden. In this study, primers were designed to identify the factor V mutation by allele-specific PCR amplification. 126 patients with thromboembolic events were analysed using this technique, PCR-RFLP and direct sequencing. The concordance between these techniques was 100%. In 27 patients a heterozygous factor VGln506 mutation was detected, whereas one patient with recurrent thromboembolism was homozygous for the point mutation. Due to its time- and cost-saving features allele-specific amplification should be considered for screening of factor VGln506.

scholarly journals WASP: a Web-based Allele-Specific PCR assay designing tool for detecting SNPs and mutations

BMC Genomics ◽  
2007 ◽  
Vol 8 (1) ◽  
pp. 275 ◽  
Author(s):  
Pongsakorn Wangkumhang ◽  
Kridsadakorn Chaichoompu ◽  
Chumpol Ngamphiw ◽  
Uttapong Ruangrit ◽  
Juntima Chanprasert ◽  
...  
Keyword(s):  
Pcr Assay ◽  
Web Based ◽  
Specific Pcr ◽  
Allele Specific ◽  
Specific Pcr Assay ◽  
Allele Specific Pcr

scholarly journals Single-Tube Genotyping without Oligonucleotide Probes

1999 ◽  
Vol 9 (1) ◽  
pp. 72-78 ◽  
Author(s):  
Søren Germer ◽  
Russell Higuchi
Keyword(s):  
Pcr Amplification ◽  
Oligonucleotide Probes ◽  
Nucleotide Polymorphisms ◽  
Single Tube ◽  
Melting Temperatures ◽  
Specific Pcr ◽  
Allele Specific ◽  
Reverse Primer ◽  
Common Reverse Primer ◽  
Template Dna

We report the development of a self-contained (homogeneous), single-tube assay for the genotyping of single-nucleotide polymorphisms (SNPs), which does not rely on fluorescent oligonucleotide probes. The method, which we call Tm-shift genotyping, combines allele-specific PCR with the discrimination between amplification products by their melting temperatures (Tm). Two distinct forward primers, each of which contains a 3′-terminal base that corresponds to one of the two SNP allelic variants, are combined with a common reverse primer in a single-tube reaction. A GC-tail is attached to one of the forward allele-specific primers to increase theTm of the amplification product from the corresponding allele. PCR amplification, Tmanalysis, and allele determination of genomic template DNA are carried out on a fluorescence-detecting thermocycler with a dye that fluoresces when bound to dsDNA. We demonstrate the accuracy and reliability ofTm-shift genotyping on 100 samples typed for two SNPs, and recommend it both as a simple and inexpensive diagnostic tool for genotyping medically relevant SNPs and as a high-throughput SNP genotyping method for gene mapping.

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