Non Gel Based Assays of PCR Porducts for SNP Detection, Amplification Due to Mismatch, Invasive Cleavage Assay, Non gel Based Approaches

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Home >> Biotechnology and Genomics >> DNA-Based Molecular Markers in Biotechnology >> Non Gel Based Assay of PCR Products for SNP Detection

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Non-gel based assays of PCR products for SNP detection.

Several non-gel based methods are avialable for SNP detection. If SNP is present at 3' end of a PCR primer binding site, it can be detected simply by the failure of amplification due to mismatch between the primer sequence and the binding site in the template, although it may be difficult to distinguish this failure of PCR due to SNP from a PCR failure due to other reasons.
Other non-gel based approaches, that are available for the detection of SNPs at an internal position of an amplicon generally need initial PCR amplification, followed by a non-gel based assay, that discriminates between wild and mutant alleles.

There are still other assays available now, which neither need separation on the gel, nor do they need PCR amplification

(e.g. 'invasive cleavage assay; for details. consult Gupta. 2001, Curr Science, Feb 26, 2001).

The common non-gel based assays for detection of SNPs at the internal sites are based on the detection of mismatch between the PCR product and an oligonucleotide used as a probe.
Following are some examples of such assays:
(a) TAqman Assay,
(b) Use of Molecular Beacon,
(c) Oligonucleotide Ligation Assay
(d) DNA chip/microarray,
(e) Dynamic Allele Specific Hybridization,
(f) Minisequencing,
(g) Pyrosequencing for SNP genotyping,
(h) Temperature modulated heteroduplex analysis using dHPLC WAVE Tm system.

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