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

How will it impact the rest of the organs, Starting at the esophagus, trace...

Starting at the esophagus, trace the pathway of food through the system. At each organ, indicate anatomical adaptations to the general GI tract tube structure that enhance the spec

What is meant by mutualist exploration, What is meant by "mutualist explora...

What is meant by "mutualist exploration of cellulose digestion", a phenomenon that occurs in some mammals and insects? Herbivorous animals eat great amounts of cellulose, a sub

Explain about the forces acting on a gravity retaining wall, Explain about ...

Explain about the forces acting on a gravity retaining wall. Forces Acting on a Gravity Retaining Wall: In a general way the forces that act on a gravity retaining wall. The

What is posterior aneurysm? , What is  Posterior Aneurysm? The te...

What is  Posterior Aneurysm? The technique of Posterior Aneurysm operation is the same. Care must be taken to avoid injury to the posterior papillary muscle and poster

Explain atrial pressure, Q. Explain Atrial Pressure? The RA pressure wa...

Q. Explain Atrial Pressure? The RA pressure wave form has three positive deflections - "a", "c", and "v" waves. The "a" wave is due to atrial systole and follows the P-wave on

Explain the use of enzyme assay, Explain the use of Enzyme assay Enzym...

Explain the use of Enzyme assay Enzyme assay is also used for research into such processes as the browning of plant products which poses problems during value addition. The br

Surface Processes on Earth, Which of the following is not a type of mass mo...

Which of the following is not a type of mass movement that results from the force of gravity? A. Landslide B. Creep C. Deflation D. Mudflow

Explain about the commercial sterilization, Explain about the Commercial St...

Explain about the Commercial Sterilization? Sterilization is the most extreme heat treatment given in preservation of food by heat. Usually target organism is a heat resistant

Animal genetics and breeding, POULTRY BREEDING Poultry breeding has acq...

POULTRY BREEDING Poultry breeding has acquired pre-eminence over all other domesticated livestock enterprises. Production level of layer and broiler type of chicken touched spe

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