scholarly journals Closed-tube genotyping of apolipoprotein E by isolated-probe PCR with multiple unlabeled probes and high-resolution DNA melting analysis

BioTechniques ◽  
2007 ◽  
Vol 43 (1) ◽  
pp. 87-91 ◽  
Author(s):  
Matthew Dean Poulson ◽  
Carl T. Wittwer
Keyword(s):  
High Resolution ◽  
Apolipoprotein E ◽  
Dna Melting ◽  
Closed Tube ◽  
Melting Analysis

High resolution DNA melting analysis: an application for prenatal control of α-thalassemia

2010 ◽  
pp. n/a-n/a
Author(s):  
Supatra Sirichotiyakul ◽  
Chanane Wanapirak ◽  
Rattika Saetung ◽  
Torpong Sanguansermsri
Keyword(s):  
High Resolution ◽  
Dna Melting ◽  
Melting Analysis

Simultaneous mutation scanning and genotyping by high-resolution DNA melting analysis

2007 ◽  
Vol 2 (1) ◽  
pp. 59-66 ◽  
Author(s):  
Jesse Montgomery ◽  
Carl T Wittwer ◽  
Robert Palais ◽  
Luming Zhou
Keyword(s):  
High Resolution ◽  
Dna Melting ◽  
Mutation Scanning ◽  
Melting Analysis

scholarly journals High-throughput genotyping of mannose-binding lectin variants using high-resolution DNA-melting analysis

2010 ◽  
Vol 31 (4) ◽  
pp. E1286-E1293 ◽  
Author(s):  
Rolf H.A.M. Vossen ◽  
Martine van Duijn ◽  
Mohamed R. Daha ◽  
Johan T. den Dunnen ◽  
Anja Roos
Keyword(s):  
High Resolution ◽  
High Throughput ◽  
Mannose Binding Lectin ◽  
Dna Melting ◽  
Mannose Binding ◽  
Melting Analysis ◽  
Binding Lectin

Validation of high-resolution DNA melting analysis for mutation scanning of the LMNA gene

2009 ◽  
Vol 42 (9) ◽  
pp. 892-898 ◽  
Author(s):  
Gilles Millat ◽  
Valérie Chanavat ◽  
Sophie Julia ◽  
Hervé Crehalet ◽  
Patrice Bouvagnet ◽  
...  
Keyword(s):  
High Resolution ◽  
Dna Melting ◽  
Lmna Gene ◽  
Mutation Scanning ◽  
Melting Analysis

31-OR: HLA matching by high-resolution DNA melting analysis for living related transplantation

2007 ◽  
Vol 68 (1) ◽  
pp. S116
Author(s):  
Luming Zhou ◽  
Hongzhe Lu ◽  
Alexander Myers ◽  
Michelle Taylor ◽  
Tom Fuller ◽  
...  
Keyword(s):  
High Resolution ◽  
Dna Melting ◽  
Hla Matching ◽  
Melting Analysis ◽  
Living Related

Development of High Resolution DNA Melting Analysis for Simultaneous Detection of Potato Mop-Top Virus and Its Vector, Spongospora subterranea, in Soil

Plant Disease ◽  
2020 ◽  
Author(s):  
Xianzhou Nie ◽  
Mathuresh Singh ◽  
Dahu Chen ◽  
Cassandra Gilchrist ◽  
Yasmine Soqrat ◽  
...  
Keyword(s):  
High Resolution ◽  
Simultaneous Detection ◽  
Soil Samples ◽  
Dna Melting ◽  
Spongospora Subterranea ◽  
Rt Pcr ◽  
Efficient Detection ◽  
Melting Analysis ◽  
Potato Mop Top Virus ◽  
Negative Detection

In this study, a set of duplex reverse transcription (RT)-PCR-mediated high resolution DNA melting (HRM) analyses for simultaneous detection of potato mop-virus (PMTV) and its protist vector, Spongospora subterranea f.sp. subterranea (Sss), was developed. The infestation of soil by PMTV was detected by using a tobacco-based baiting system. Total RNA extracted from the soil led to successful RT-PCR gel-electrophoresis detection of both PMTV and Sss. To facilitate more efficient detection, newly designed primer pairs for PMTV RNA species (i.e., RNA-Rep, -CP, and -TGB) were analyzed together with the existing Sss primers using real-time RT-PCR. The resulting amplicons exhibited melting profiles that could be readily differentiated. Under duplex RT-PCR format, all PMTV and Sss primer combinations led to successful detection of respective PMTV RNA species and Sss in the samples by high resolution DNA melting (HRM) analyses. When the duplex HRM assay was applied to soil samples collected from six fields at four different sites in New Brunswick, Canada, positive detection of PMTV and/or Sss was found in 63-100% samples collected from fields in which PMTV-infected tubers had been observed. In contrast, the samples from fields where neither PMTV- nor Sss-infected tubers had been observed resulted in negative detection by the assay. Bait tobacco bioassay for PMTV and Sss produced similar results. Between 63%-83% and 100% of the soil samples collected from PMTV-infested fields led to PMTV and Sss infections in the bait tobacco plants, respectively; whereas no PMTV or Sss infected plants were obtained from soil samples collected from PMTV/Sss-free fields.

scholarly journals High-Resolution Genotyping by Amplicon Melting Analysis Using LCGreen

2003 ◽  
Vol 49 (6) ◽  
pp. 853-860 ◽  
Author(s):  
Carl T Wittwer ◽  
Gudrun H Reed ◽  
Cameron N Gundry ◽  
Joshua G Vandersteen ◽  
Robert J Pryor
Keyword(s):  
High Resolution ◽  
Mutation Screening ◽  
Melting Curve ◽  
Single Nucleotide ◽  
Mutation Scanning ◽  
Double Stranded Dna ◽  
Closed Tube ◽  
Pcr Products ◽  
Melting Analysis ◽  
Detection Of Mutations

Abstract Background: High-resolution amplicon melting analysis was recently introduced as a closed-tube method for genotyping and mutation scanning (Gundry et al. Clin Chem 2003;49:396–406). The technique required a fluorescently labeled primer and was limited to the detection of mutations residing in the melting domain of the labeled primer. Our aim was to develop a closed-tube system for genotyping and mutation scanning that did not require labeled oligonucleotides. Methods: We studied polymorphisms in the hydroxytryptamine receptor 2A (HTR2A) gene (T102C), β-globin (hemoglobins S and C) gene, and cystic fibrosis (F508del, F508C, I507del) gene. PCR was performed in the presence of the double-stranded DNA dye LCGreen, and high-resolution amplicon melting curves were obtained. After fluorescence normalization, temperature adjustment, and/or difference analysis, sequence alterations were distinguished by curve shape and/or position. Heterozygous DNA was identified by the low-temperature melting of heteroduplexes not observed with other dyes commonly used in real-time PCR. Results: The six common β-globin genotypes (AA, AS, AC, SS, CC, and SC) were all distinguished in a 110-bp amplicon. The HTR2A single-nucleotide polymorphism was genotyped in a 544-bp fragment that split into two melting domains. Because melting curve acquisition required only 1–2 min, amplification and analysis were achieved in 10–20 min with rapid cycling conditions. Conclusions: High-resolution melting analysis of PCR products amplified in the presence of LCGreen can identify both heterozygous and homozygous sequence variants. The technique requires only the usual unlabeled primers and a generic double-stranded DNA dye added before PCR for amplicon genotyping, and is a promising method for mutation screening.

Validation of high-resolution DNA melting analysis for mutation scanning of the CDKL5 gene: Identification of novel mutations

Gene ◽  
2013 ◽  
Vol 512 (1) ◽  
pp. 70-75 ◽  
Author(s):  
Laure Raymond ◽  
Bertrand Diebold ◽  
Céline Leroux ◽  
Hélène Maurey ◽  
Valérie Drouin-Garraud ◽  
...  
Keyword(s):  
High Resolution ◽  
Gene Identification ◽  
Dna Melting ◽  
Novel Mutations ◽  
Mutation Scanning ◽  
Melting Analysis
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