Question

Can rhamp genotyping for multiple gene do in single tube

Answer 1

The description of the polymerase chain reaction in 1985 caused a revolution in genetics and today molecular diagnostics is one of the leading growth areas across all disciplines of laboratory medicine. This paper reviews the principles and limitations of a number of traditional and emerging techniques for typing of single nucleotide substitutions. The techniques discussed include traditional approaches such as restriction enzyme analysis, more recent homogenous methods, such as those utilising TaqMan®, fluorescence resonance energy transfer (FRET) and Scorpion® probes, and high resolution melting curve analysis. Non-homogenous but highly flexible approaches such as Pyrosequencing™ and mass-spectrometry are also discussed. While many techniques are available, it is clear that no one approach is clearly superior. However, in terms of their many advantages and continuing developments, homogenous approaches have much to recommend them.Allele-Specific Amplification

Allele-Specific AmplificationAllele-specific amplification, also known as the amplification refractory mutation system (ARMS), uses allele specific oligonucleotide (ASO) PCR primers and was an early and commonly used PCR based method for genotyping.3 In this approach, one of the two oligonucleotide primers used for PCR is designed to bind to the mutation site, most commonly with the 3' end of the primer targeting the mutation site. Under carefully controlled conditions (annealing temperature, magnesium concentration etc.),

Ligation Based Assays

Ligation Based AssaysA number of approaches make use of DNA ligase, an enzyme that can join two adjacent oligonucleotides hybridised to a DNA template. The specificity of the approach comes from the requirement for a perfect match between the hybridised oligonucleotides and the DNA template at the ligation site. In the oligonucleotide ligation assay

Answer 2

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