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 the acquired deficiency of the vitamin k metabolism, Why are some t...

Why are some types of hemorrhagic diseases caused by genetic or acquired deficiency of the vitamin K metabolism? Lack of vitamin K predisposes to hemorrhages as this vitamin i

LIVING THINGS, CHARACTERICTICS OF PLATYHELMENTUS

CHARACTERICTICS OF PLATYHELMENTUS

Name the phases into which hiv infection is often divided, What are the thr...

What are the three phases into which the HIV infection is often divided? The HIV infection is often separated into three phases: the acute phase, from the infection to 1 until

What is gene, What is gene? Gene is a sequence of DNA nucleotides that ...

What is gene? Gene is a sequence of DNA nucleotides that codifies the production of a protein.

Explain theory of enzyme action, Explain Theory of enzyme action L. Mic...

Explain Theory of enzyme action L. Michaelis  and k L. Menten developed a general  theory of enzyme action and kinetics  in  1913. According to this theory, we learnt  that

What are the endocrine functions of the placenta, What are the endocrine fu...

What are the endocrine functions of the placenta? The placenta has endocrine function since it secretes the hormones progesterone and estrogen that maintain the endometrium (in

Explain strengthening the public health measures - anaemia, Explain Strengt...

Explain Strengthening the public health measures? Parasitic infestations and protozoal infections cause iron deficiency through loss of blood or destruction of red blood cells.

Modern theory in biology, Modern Theory Naturalistic theory: Accordin...

Modern Theory Naturalistic theory: According to this theory Life originated upon our earth spontaneously from nonliving matter. But there are two significant points in this r

How are ecological interactions classified, How are ecological interactions...

How are ecological interactions classified? Ecological interactions are divided as intraspecific or interspecific interactions and as harmonious or inharmonious interactions.

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