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

What is metabolomics, Together, genomics, proteomics and transcriptomics ar...

Together, genomics, proteomics and transcriptomics are influential approaches to enhancing our understanding not only of how cells function normally but also what the key modificat

Pathophysiology of aortic regurgitation, Q. Pathophysiology of aortic regur...

Q. Pathophysiology of aortic regurgitation? Left ventricle responds to chronic aortic regurgitation by chamber dilatation and an increase in its compliance so that end diastoli

Why dietary supplements do not speed up a child''s growth, Why Dietary supp...

Why Dietary supplements do not speed up a child's growth? Dietary supplements do not "speed up" a child's growth and development: There is no scientific evidence that me

What is sustainability, What is sustainability? Sustainability is the a...

What is sustainability? Sustainability is the ability to meet human needsin a way that will allow a human population to survive indefinitely.

Structure and fate of endosperm, Structure and Fate of Endosperm The ...

Structure and Fate of Endosperm The cells of endosperm are usually thin-walled, large isodiametric and devoid of pits and store large amount of food materials. The starch alo

Locomotion in mollusca, Locomotion in Mollusca The major locomotor or...

Locomotion in Mollusca The major locomotor organ in Mollusca is the foot, which is a characteristic feature of these animals. In its simplest form the foot is a flit ventral

Respiration, abnormal lung volumes and capacities

abnormal lung volumes and capacities

Ecology, Importance of ecology

Importance of ecology

What is the use of uncoated polyvinyl chloride, Q. What is the use of Uncoa...

Q. What is the use of Uncoated polyvinyl chloride? Uncoated polyvinyl chloride (PVC) and polyvinylidene chloride (PVDC): PVC and PVDC are closely related plastics that are a

Ecosystems, what is biodiversity?where are ecosystems with the highest biov...

what is biodiversity?where are ecosystems with the highest bioversityfound?

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