ReviewFrontiers in molecular biosciences2022
Designing Allele-Specific Competitive-Extension PCR-Based Assays for High-Throughput Genotyping and Gene Characterization.
Review in Frontiers in molecular biosciences, 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers, 1 of them a synthesis that pooled it.
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Who cites it
20 citing papers in PubMed, 1 synthesis or guideline pooled it.
- A plethora of laboratory protocols for vitamin D receptor (VDR) gene variants detection: a systematic review of associations with hypertensive disorders of pregnancy.BMC pregnancy and childbirth · 2025Pooled it
- Detection and Characterization of Salmonella enterica Serotypes by Simple PCR Technologies.Methods in molecular biology (Clifton, N.J.) · 2027Article
- GWAS-Guided Compact SNP Panels Enable Breeding-Relevant Prediction of Bolting and Flowering Timing of Lettuce.Plants (Basel, Switzerland) · 2026Article
- Development and Application of KASP Markers for Candidate Glucosinolate Biosynthesis Genes in Broccoli.International journal of molecular sciences · 2026Article
- Article
- A Species-SpecificBiology · 2026Article
- A Comparative Review of Methods for Detecting Epidermal Growth Factor Receptor Mutations in Cell-free DNA from Lung Cancer Patients.Current molecular medicine · 2026Review
- From sequencing to understanding: a grand challenge in genome-scale molecular and genetic analysis.Frontiers in plant science · 2026Article
- EasyKASP: a simple and fast tool for KASP primer design.BMC bioinformatics · 2025Article
- Comprehensive web-based platform for advanced PCR design, genotyping, synthetic biology, molecular diagnostics, and sequence analysis.Molecular therapy. Nucleic acids · 2025Article
- Determining Zygosity With Multiplex Kompetitive Allele-Specific PCR (mxKASP) Genotyping.Ecology and evolution · 2025Article
- An Assay for Variable Fragment Length Allele-Specific Genotyping.Methods in molecular biology (Clifton, N.J.) · 2025Article
- PCR Allele Competitive Extension (PACE) Workflow for Genotyping by SNP Identification.Methods in molecular biology (Clifton, N.J.) · 2025Article
- [Progression and application of circulating tumor DNA in lymphoma].Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi · 2024Review
- Genotyping single point mutations in rd1 and rd8 mice using melting curve analysis of qPCR fragments.Scientific reports · 2024Article
- The Kirkhouse Trust: Successes and Challenges in Twenty Years of Supporting Independent, Contemporary Grain Legume Breeding Projects in India and African Countries.Plants (Basel, Switzerland) · 2024Review
- Integrating Artificial Intelligence for Advancing Multiple-Cancer Early Detection via Serum Biomarkers: A Narrative Review.Cancers · 2024Review
- Article
- CRISPR/Cas9-gene editing approaches in plant breeding.GM crops & food · 2023Review
- Editorial: Spectroscopy for crop and product phenotyping.Frontiers in plant science · 2022Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Polymerase chain reaction (PCR) is a simple and rapid method that can detect nucleotide polymorphisms and sequence variation in basic research applications, agriculture, and medicine. Variants of PCR, collectively known as allele-specific PCR (AS-PCR), use a competitive reaction in the presence of allele-specific primers to preferentially amplify only certain alleles. This method, originally named by its developers as Kompetitive Allele Specific PCR (KASP), is an AS-PCR variant adapted for fluorescence-based detection of amplification results. We developed a bioinformatic tool for designing probe sequences for PCR-based genotyping assays. Probe sequences are designed in both directions, and both single nucleotide polymorphisms (SNPs) and insertion-deletions (InDels) may be targeted. In addition, the tool allows discrimination of up to four-allelic variants at a single SNP site. To increase both the reaction specificity and the discriminative power of SNP genotyping, each allele-specific primer is designed such that the penultimate base before the primer's 3' end base is positioned at the SNP site. The tool allows design of custom FRET cassette reporter systems for fluorescence-based assays. FastPCR is a user-friendly and powerful Java-based software that is freely available (http://primerdigital.com/tools/). Using the FastPCR environment and the tool for designing AS-PCR provides unparalleled flexibility for developing genotyping assays and specific and sensitive diagnostic PCR-based tests, which translates into a greater likelihood of research success.
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