Enzymatic mutation detection method, Biology

Assignment Help:

This technique utilizes an enzyme resolvase, endo vii, cloned from the bacteriophage t4. This enzyme has high specificity to find deletions, insertions, and base substitutions mutations or any mismatches. First the pcr is utilized to intensify the normal and mutant allele of targeted sequence, these pcr primers are labeled with fam(6-carboxyfluoroscein amidite) which is a blue fluorescent dye and reverse primer with tet(4,7,2,7-tetra chloro-6-carboxy fluoroscein) amidite which is a green fluorescent dye.

Denaturation and renaturation of normal and mutant allele in a mixture is made, consequently mismatch heteroduplexes will form about 50 % of the time, each and every base pair creates two mismatch. The endo vii enzymes then scan the double stranded dna until it finds structural distortion, either a bubble made by single base pair mutations or a hetero duplex loop created by hybridization of a wild type allele with a mutant allele having an insertion or deletion. The enzyme cleaves with in 6 base pair on 3"side of mutation creating two smaller fragments, one blue and the other one green. The dna sequence is analyzed on automated dna sequence and mobility of every fragment is analyzed.

 


Related Discussions:- Enzymatic mutation detection method

Difference between adrenergic and cholinergic fibre, Difference between Adr...

Difference between Adrenergic and Cholinergic Fibre- S.NO .   1. ADRENERGI C FIBRE   Release nor adrenaline at their CHOLINE

What is the evolutionary importance of pteridophytes, What is the evolution...

What is the evolutionary importance of pteridophytes? As the first tracheophytes, pteridophytes were also the first plants to extensively colonize the terrestrial environment f

Muromonab cd-3 , Muromonab CD-3 (Orthoclone, OKT-3) A monoclonal anti...

Muromonab CD-3 (Orthoclone, OKT-3) A monoclonal antibody against mature T lymphocytes, OKT-3 is primarily used to treat acute rejection episode. IV for 10-14 days. Effe

Explain asystole and pulseless electrical activity, Explain Asystole and Pu...

Explain Asystole and Pulseless Electrical Activity (PEA) The outcomes from these rhythms are generally much worse than for VT/VF, unless a readily correctable cause is identif

What is genomics , We are living in an unprecedented age of biological ...

We are living in an unprecedented age of biological discovery and the application of biological knowledge.   Programmed DNA sequencing delivered, in the year of  2001 over the 2.6

How ignorance and poor socioeconomic cause pem, How Ignorance and Poor Soci...

How Ignorance and Poor Socioeconomic cause PEM? Status Improper childcare, either as a result of lack of knowledge or lack of time for mother, could also contribute to the onse

What does vacuoles do, What does vacuoles do? It is an organelle, locat...

What does vacuoles do? It is an organelle, located in the cell's cytoplasm, which is the site of protein synthesis. It is really a protein having of 2 parts. These 2 "parts"

Explain about ecological species concept, Q. Explain about Ecological speci...

Q. Explain about Ecological species concept? The ecological species concept: This identifies a species as a population that occupies a distinct ecological niche, whence its rep

Determine the function of osseointegration, Determine the function of Osseo...

Determine the function of Osseointegration Osseointegration is also a measure of implant stability, which occurs at two stages- primary and secondary. Primary stability has be

Write Your Message!

Captcha
Free Assignment Quote

Assured A++ Grade

Get guaranteed satisfaction & time on delivery in every assignment order you paid with us! We ensure premium quality solution document along with free turntin report!

All rights reserved! Copyrights ©2019-2020 ExpertsMind IT Educational Pvt Ltd