A modified simple RFLP-PCR method for single nucleotide polymorphism (SNP) typing
نویسندگان
چکیده
We describe a modified single nucleotide polymorphism (SNP) typing method based on the restriction fragment length polymorphism polymerase chain reaction (RFLP-PCR). This is a simple, economical method without the need for special equipment. For most SNP loci, a common restriction endonuclease (Hind III, EcoR I or BamH I) recognizing site (RER) can be introduced into one allelic form, but not the other by two rounds of mismatched PCR. The flanking regions can be changed by as many as five bases after PCR amplification with specially designed mismatching primers so the genotypes can be distinguished after digestion of the PCR products with corresponding endonucleases.
منابع مشابه
A Simplified Van Erth Single Nucleotide Polymorphism (SNP) Typing Method of Bacillus Anthracis Applicable by Traditional Thermocycler Machines
Abstract SNP typing is now a well-established genotyping system in Bacillus anthracis studies. In the original standard method of Van Erth, SNPs at 13 loci of the B. anthracis genome were analyzed. In order to simplify and make appropriate this expensive method to low-budget laboratory settings, 13 primer pairs targeting the 13 corresponding SNPs were designed. Besides, a universal PCR proto...
متن کاملSNP Cutter: a comprehensive tool for SNP PCR–RFLP assay design
The Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is a relatively simple and inexpensive method for genotyping single nucleotide polymorphisms (SNPs). It requires minimal investment in instrumentation. Here, we describe a web application, 'SNP Cutter,' which designs PCR-RFLP assays on a batch of SNPs from the human genome. NCBI dbSNP rs IDs or formatted SNPs are ...
متن کاملP-196: Association rs3819392 Single Nucleotide Polymorphism within The KIT Gene with Azoospermic Male Infertility
Background: Recent studies have shown that KIT is expressed in the cytoplasm of the spermatogonia, acrosomal granules and leydig cells. Reduction in KIT expression in oligozoospermia with an increase in the germ cell apoptosis process. Three single-nucleotide polymorphisms (SNPs) have been identified and these have been studied to discover KIT role in the male infertility. The aim of this study...
متن کاملGenotyping of polymorphisms in alcohol and aldehyde dehydrogenase genes by direct application of PCR-RFLP on dried blood without DNA extraction.
We have developed a simple, labor-saving, inexpensive, and rapid single nucleotide polymorphism (SNP) genotyping method that works directly on whole human blood. This single-tube genotyping method was used to successfully and reliably genotype ADH1B and ALDH2 polymorphisms without DNA isolation using a 1.2-mm disc of dried blood and the KOD FX PCR enzyme kit. SNP genotyping was performed by a p...
متن کاملNatural SNP-RFLP Primer Design using Memetic Algorithm
Single nucleotide polymorphisms (SNPs) can be genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Before performing PCR-RFLP for SNP genotyping, a feasible primer pair observes numerous constraints and an available restriction enzyme discriminate the target SNP, are required, here we called it natural SNP-RFLP primer design. Design the natural SNP-RFLP pr...
متن کامل