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

Explain interaction of pyridoxine with carbohydrates, Explain Interaction o...

Explain Interaction of pyridoxine with Carbohydrates? Pyridoxine is involved in glyconeogenesis through its action in transaminase reactions. Low levels of pyridoxine impair gl

Of what importance are lactic acid fermentation to the cell, Of what import...

Of what importance are lactic acid fermentation and alcoholic fermentation to the cells that use these pathways? These pathways regenerate NAD, which the cells can use to keep

Management of diet for diabetes, Q. Management of Diet for diabetes? Di...

Q. Management of Diet for diabetes? Diet plays a very important role in management of diabetes as it exerts a direct influence on the blood glucose levels. It is one of the vit

Reproductive isolation, REPRODUCTIV E ISOLATION - According to the bio...

REPRODUCTIV E ISOLATION - According to the biological species concept, members of one species are reproductively isolated from members of all other species. Accordingly, re

How can you get a replacement, The book that I rented is missing some pages...

The book that I rented is missing some pages. How can i get a replacement?

How long is the incubation period of the hiv, Q. How long is the incubation...

Q. How long is the incubation period of the HIV? What is meant by acute AIDS? The incubation period of the HIV that the time interval between the infection and the beginning of

Define measuring body composition - underwater weighing, Define Techniques ...

Define Techniques for Measuring Body Composition - Underwater weighing? Underwater weighing (densitometry): It applies the Archimedean principle of water displacement by the fu

Explain sound production - ethological approach in taxonomy, Explain Sound ...

Explain Sound production - Ethological Approach in Taxonomy The most used, ethological feature in identification is the sound produced by various species of animals. This is

Explain treatment of acute pericarditis, Q. Explain Treatment of Acute peri...

Q. Explain Treatment of Acute pericarditis? Symptomatic Treatment Pain relief can be achieved with indomethacin 25 to 50 mg. thrice daily; Ibuprofen 600 mg. thrice daily

Green glands of crustaceans, Normal 0 false false false ...

Normal 0 false false false EN-IN X-NONE X-NONE MicrosoftInternetExplorer4

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