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 are the possible genotypes for its parents, Roan (RW) is a color of ca...

Roan (RW) is a color of cattle in which both red and white hairs are present due to codominance. What are the phenotype and genotype ratios of offspring produced by: roan bull and

Signify gene interaction effects, The type of gene interaction in which eff...

The type of gene interaction in which effects of one gene override or mask the effects of other entirely different genes is known as: a) Mutation b) Linkage c) Pleitropy

Explain milling extraction rate, Explain Milling extraction rate The ch...

Explain Milling extraction rate The chemical composition of the flour depends on the milling extraction rate.  Increasing the rate of flour extraction decreases the proportion

Gregor mendel''s law of independent segregation, Gregor Mendel's Law of Ind...

Gregor Mendel's Law of Independent Segregation referred to which of the following genetic elements? A. Alleles of two genes that reside on the similar chromosome B. Alleles

Poultry and duck diseases-avian encephalomyelitis, Avian encephalomyelitis ...

Avian encephalomyelitis This is an acute infectious disease of birds (3 weeks old), which is caused by a member of the genus Enterovirus in the family Picornaviridae. Epid

What is class polychetae - marine worms, What is Class Polychetae - Marine ...

What is Class Polychetae - Marine Worms? The polychaetes differ from the oligochaetes in several ways. The name itself suggests one of the differences. The prefix "poly" means

Spermatocytogenesis, SPERM A T OCY T OGENESIS In this process four...

SPERM A T OCY T OGENESIS In this process four spermatid develop from one PGC. (i) Multiplication phase The spermotogonia or sperm mother cells lie next to the b

Determine the intracellular concentration of trva, Determine the intracellu...

Determine the intracellular concentration of TRVA Consider Neuron B in the frog central nervous system whose plasma membrane has a previously unknown channel that is selectly c

Arterial blood gas studies, Arterial Blood Gas Studies: Purpose 1)  A me...

Arterial Blood Gas Studies: Purpose 1)  A measurement of partial pressure of oxygen and carbon dioxide  in arterial blood, as well as the pH  of  the blood.  2) The Partial p

What is the large size of cephalopods, Q How is the large size of some ceph...

Q How is the large size of some cephalopods related to the kind of circulatory system they present? In cephalopods the circulatory system is closed and this provides more press

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