What is allele specific PCR?

Allele-specific polymerase chain reaction (AS-PCR) is a technique based on allele-specific primers, which can be used to analyze single nucleotide polymorphism (SNP) effectively including the transition, transversion and insertion/deletion polymorphism and has been exploited in the study of diseases research, molecular …

How does allele specific PCR work?

The Allele-specific PCR has the power to detect a single specific allele. Meaning, If you wish to amplify only a mutant allele, design a primer set accordingly and amplify it using this technique. Each set of specific primers is designed for each specific allele.

What are the potential applications of allele specific PCR?

Potential applications include gene mapping, genetic disease diagnosis, HLA typing, paternity testing, and forensic science.

What is allele specific amplification?

Allele specific amplification differs from the normal PCR in that primers are carefully designed to target the region of difference in the different alleles so that a pair of primers only amplify a specific allele but not others.

What is allele specific primer?

Allele Specific Primer Extension (ASPE) is a solution based, sequence specific enzymatic reaction technology that can be used to assay multiple SNPs in a single tube. This is done with the help of an appropriate capture sequence attached to the allele specific oligonucleotide.

What is multiplex PCR used for?

Multiplex PCR is used in life science research, clinical diagnostics, and forensic laboratories. The development of PCR detection systems with simultaneous multi-target detection and advances in probe chemistries have made comparative analyses standard in many areas of research and testing.

What are the 3 steps in PCR?

PCR is based on three simple steps required for any DNA synthesis reaction: (1) denaturation of the template into single strands; (2) annealing of primers to each original strand for new strand synthesis; and (3) extension of the new DNA strands from the primers.

How does an allele specific qPCR work?

The allele-specific qPCR (ASQ) does not require post-PCR processing and can genotype germline mutants through either threshold cycle (C t) or end-point fluorescence reading. ASQ utilizes allele-specific primers, a locus-specific reverse primer, universal fluorescent probes and quenchers, and hot start DNA polymerase.

Which is better RFLP or allele specific PCR?

Although simpler than RFLP, current versions of allele-specific PCR may still require post-polymerase chain reaction (PCR) handling such as sequencing, or they are more expensive if allele-specific fluorescent probes are used.

How are allele specific primers used in ASQ?

ASQ utilizes allele-specific primers, a locus-specific reverse primer, universal fluorescent probes and quenchers, and hot start DNA polymerase. Individual laboratories can further optimize this open-source system as we completely disclose the sequences, reagents, and thermal cycling protocol.

How is quantitative PCR used for genotyping?

Therefore, we developed a one-step open-source genotyping method based on quantitative PCR. The allele-specific qPCR (ASQ) does not require post-PCR processing and can genotype germline mutants through either threshold cycle (C t) or end-point fluorescence reading.