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 polymerase chain reaction (AS-PCR), also known as amplification refractory mutation system (ARMS) or PCR amplification of specific alleles (PASA) is a PCR-based method which can be employed to detect the known SNPs [7]. The concept of AS-PCR was initiated by Newton et al.
What is the principle behind PCR?
Polymerase chain reaction (PCR) is a technology used for quick and easy amplifying DNA sequences, which is based on the principle of enzymatic replication of the nucleic acids. This method has in the field of molecular biology an irreplaceable role and constitutes one of the basic methods for DNA analysis.
What is the principle of allele specific PCR?
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 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.
Why is PCR important in biotechnology?
PCR technique gives researchers the means to make more DNA by synthesising multiple copies of specific DNA fragments using DNA polymerase.
What is the principle of Methylation Specific PCR?
Methylation-specific PCR (MS-PCR or MSP) is one of the most commonly used methods for gene/sequence-specific detection of DNA methylation. The DNA undergoes bisulfite conversion of cytosine to uracil and then the methylated sequences are selectively amplified with primers specific for methylation.
Can PCR detect DNA methylation?
Methylation specific PCR (Polymerase Chain Reaction) is used to detect genes or sequences with DNA methylation.
What is Dcaps marker?
assay is a modification of CAPS (or alternatively, PCR-RFLP) technique for detection of Single Nucleotide Polymorphisms (SNPs).
How does the allele-specific PCR principle 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.
Where is the allele specific nucleotide in a primer?
When designing your primers, the allele-specific nucleotide should ideally be the last, or at least one of the last two nucleotides at the 3′ end of the primer. If you still have specificity problems after making these changes, you might want to try primers with an additional intentional mismatch.
How did the concept of as-PCR come about?
The concept of AS-PCR was initiated by Newton et al. [ 8 ], approximately six years after PCR was invented. In this approach, the specific primers are designed to permit amplification by DNA polymerase only if the nucleotide at the 3’-end of the primer perfectly complements the base at the variant or wild-type sequences.
How is PCR used to detect mutations in DNA?
The PCR technique can detect mutations like deletion, duplications, insertion or single base change- SNP. It’s a temperature-dependent amplification technique that relies on Taq DNA polymerase . Many variations of PCR exist depending upon the assay requirement, ARMS-PCR is one among them.