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

Illustrate natural and phylogenetic classification, Q. Illustrate Natural a...

Q. Illustrate Natural and Phylogenetic Classification? Natural Classification Natural systems of classification reflect the situation as it might have existed in nature. T

Illustrate stage of cellular respiration, Q. What is the stage of cellular ...

Q. What is the stage of cellular respiration during which carbon dioxide is liberated? In aerobic cellular respiration the release of carbon dioxide happens in the transformati

Stretch receptors, Stretch Receptors Stretch receptors able of percept...

Stretch Receptors Stretch receptors able of perception of muscle tension or stretch are distributed in the connective tissue related with muscles of many groups of animals. Th

Explain about the imvic test, Explain about the IMViC Test? IMViC test ...

Explain about the IMViC Test? IMViC test is a combination of four tests: (1) Indole production (2) Methyl Red test (3) Voges Proskauer, and (4) Citrate Utilization

Why protein requirements of athletes get increased, Why Protein requirement...

Why Protein requirements of athletes get increased? Protein requirements of athletes, particularly those engaged in strength and power events, may be increased above those of s

Prior to the solving of the structure of dna, Prior to the solving of the s...

Prior to the solving of the structure of DNA, Erwin Chargaff calculated the amounts of the four nucleotides (A, T,G,C) and made which of the following conclusions? A. All four

Describe open circulatory system, Q. What is an open circulatory system? ...

Q. What is an open circulatory system? Open circulatory system is the one in which blood doesn't circulate only inside blood vessels but it also falls in cavities that irrigate

Define exocytosis , A cell frequently wants to secrete larger molecule...

A cell frequently wants to secrete larger molecules than can be accommodated through the transport systems dealt with in Topic E3. Exocytoses get to the movement of proteins out of

Pathophysiology, The clinical  manifestations of IE result from the ...

The clinical  manifestations of IE result from the local destructive effects of intracardiac infection; the embolization of bland or septic fragments of vegetations to

Explain about the parkinson''s disease, Explain about the Parkinson's disea...

Explain about the Parkinson's disease? Parkinson's disease is a degenerative central nervous system (CNS) condition characterized by progressive loss of cells within substantia

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