ArticleBMC genomics2009
Temperature switch PCR (TSP): Robust assay design for reliable amplification and genotyping of SNPs.
Article in BMC genomics, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Development of a KASP marker set for high-throughput genotyping in Japanese barley breeding programs with various end-use purposes.Breeding science · 2025Article
- Combination of GWAS and FMolecular genetics and genomics : MGG · 2023Article
- Quantitative Trait Loci Mapping and Identification of Candidate Genes Linked to Fruit Acidity in Apricot (Frontiers in plant science · 2022Article
- Establishment and application of a novel method based on single nucleotide polymorphism analysis for detecting β-globin gene cluster deletions.Scientific reports · 2020Article
- Development and validation of high-throughput and low-cost STARP assays for genes underpinning economically important traits in wheat.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2020Article
- Fine mapping of Rha2 in barley reveals candidate genes for resistance against cereal cyst nematode.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2019Article
- Article
- A resequencing pathogen microarray method for high-throughput molecular diagnosis of multiple etiologies associated with central nervous system infection.Archives of virology · 2017Article
- High-resolution melting combines with Bayes discriminant analysis: a novel hepatitis C virus genotyping method.Clinical and experimental medicine · 2017Article
- Gene Classification and Mining of Molecular Markers Useful in Red Clover (Frontiers in plant science · 2017Article
- Additive QTLs on three chromosomes control flowering time in woodland strawberry (Horticulture research · 2017Article
- Development of a highly sensitive real-time nested RT-PCR assay in a single closed tube for detection of enterovirus 71 in hand, foot, and mouth disease.Archives of virology · 2016Article
- Development and validation of KASP assays for genes underpinning key economic traits in bread wheat.TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik · 2016Article
- Simultaneous detection of four different neuraminidase types of avian influenza A H5 viruses by multiplex reverse transcription PCR using a GeXP analyser.Influenza and other respiratory viruses · 2016Article
- Assignment of SNP allelic configuration in polyploids using competitive allele-specific PCR: application to citrus triploid progeny.Annals of botany · 2013Article
- A high throughput multiplex PCR assay for simultaneous detection of seven aminoglycoside-resistance genes in Enterobacteriaceae.BMC microbiology · 2013Article
- Article
- A two-tube multiplex reverse transcription PCR assay for simultaneous detection of sixteen human respiratory virus types/subtypes.BioMed research international · 2013Article
- The development of a GeXP-based multiplex reverse transcription-PCR assay for simultaneous detection of sixteen human respiratory virus types/subtypes.BMC infectious diseases · 2012Article
- Simultaneously typing nine serotypes of enteroviruses associated with hand, foot, and mouth disease by a GeXP analyzer-based multiplex reverse transcription-PCR assay.Journal of clinical microbiology · 2012Article
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3 authors.
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Abstract
backgroundMany research and diagnostic applications rely upon the assay of individual single nucleotide polymorphisms (SNPs). Thus, methods to improve the speed and efficiency for single-marker SNP genotyping are highly desirable. Here, we describe the method of temperature-switch PCR (TSP), a biphasic four-primer PCR system with a universal primer design that permits amplification of the target locus in the first phase of thermal cycling before switching to the detection of the alleles. TSP can simplify assay design for a range of commonly used single-marker SNP genotyping methods, and reduce the requirement for individual assay optimization and operator expertise in the deployment of SNP assays.
resultsWe demonstrate the utility of TSP for the rapid construction of robust and convenient endpoint SNP genotyping assays based on allele-specific PCR and high resolution melt analysis by generating a total of 11,232 data points. The TSP assays were performed under standardised reaction conditions, requiring minimal optimization of individual assays. High genotyping accuracy was verified by 100% concordance of TSP genotypes in a blinded study with an independent genotyping method.
conclusionTheoretically, TSP can be directly incorporated into the design of assays for most current single-marker SNP genotyping methods. TSP provides several technological advances for single-marker SNP genotyping including simplified assay design and development, increased assay specificity and genotyping accuracy, and opportunities for assay automation. By reducing the requirement for operator expertise, TSP provides opportunities to deploy a wider range of single-marker SNP genotyping methods in the laboratory. TSP has broad applications and can be deployed in any animal and plant species.
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