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

Incubator, Incubator The incubators are essential to provide an ideal ...

Incubator The incubators are essential to provide an ideal micro environment. The main functions are: isolation, maintenance of thermo neutral ambient temperature, desired h

How gender affects the bmr, How Gender affects the bmr? We have already...

How Gender affects the bmr? We have already emphasized earlier that sex difference in metabolic rates are primarily attributable to difference in body size and composition. Wom

Human skeleton, what are the main coponents of human skeleton?

what are the main coponents of human skeleton?

Determine the genotypes of parents, In humans, the recessive allele that ca...

In humans, the recessive allele that causes a form of red-green colour blindness (c) is found on the X chromosome. a) Determine the genotypes and phenotypes of the F1 generation fr

Determine the evaluation of the bone, Evaluation of the bone Evaluation...

Evaluation of the bone Evaluation of the bone -implant interface needs to be done by the following: Failures in Implant Dentistry 1) Implant mobility and discomfort 2)

The importance of buffer systems in the human body, Outline the importance ...

Outline the importance of buffer systems in the human body. In addition, describe the main biological buffer systems and how they function.

Determine the significance of mesoglea, Determine the significance of Mesog...

Determine the significance of Mesoglea. The jellylike layer found between endodermal and ectodermal cell layers of diploblastic organisms. It acts as a type of cement holding t

Describe in detail about the cytoplasm, Describe in detail about the Cytopl...

Describe in detail about the Cytoplasm Cytoplasm also possesses a number of dense granular elements (about 25,000 per cell) called the ribosomes, which are the sites of protein

Human skeleton, HUMAN SKELETON - Hard supportive or protective elements...

HUMAN SKELETON - Hard supportive or protective elements of the animal body form the skeleton system. It's study in osteology. EXOSKELETO N - Present outside skin. It

How to investigate aortic stenosis by exercise testing, Q. How to investiga...

Q. How to investigate aortic stenosis by Exercise Testing? Exercise testing is relatively contraindicated in patients with aortic stenosis. Its diagnostic accuracy to pick up a

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