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 about the management of food allergies, Normal 0 false ...

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

Agro industrial-iodine, Iodine Iodine functions are essential componen...

Iodine Iodine functions are essential components of the thyroid hormones thyroxine (T 4) and triiodothyronine (T3), which regulate the rate of energy metabolism. Absorbed iodi

Roller pump- type of blood pump, Roller Pump: This is the type most commo...

Roller Pump: This is the type most commonly used in clinical perfusion. It has a circular metal housing inside which there is a rotating urn with two rollers fixed at 180 degrees

Define biomaterial and an implant, Define biomaterial and an implant and st...

Define biomaterial and an implant and state their role in general Biomaterial : A pharmacologically inert material that is capable of interacting with a living organism witho

Effects on ecosystem - air pollutants, Effects on Ecosystem - Air pollutant...

Effects on Ecosystem - Air pollutants The effects of pollutants on ecosystem become visible after a long period of exposure. In highly industrialised countries the vegetation

Define the classification of proteins, Define the Classification of Protein...

Define the Classification of Proteins? Proteins vary widely in. their properties. We suggest you look up this unit once again now as the information about proteins and their pr

Open and closed type of circulatory systems, Open and Closed Type of Circul...

Open and Closed Type of Circulatory Systems There are two categories of circulatory system found in higher metazoans. In one type the original blastocoel carries on to be the

Illustrate the major events of the final mitotic period, Q. What are the ma...

Q. What are the major events of the final mitotic period? The final mitotic phase is the telophase. In the telophase the following events occur decondensation of chromosomes ea

How hiv infection is mainly spread, It is mostly spread by sexual contact, ...

It is mostly spread by sexual contact, blood transfers and from HIV infected mother to child.

X linked in the calico cat, A calico female has a litter of kitten which in...

A calico female has a litter of kitten which includes several calico females a yellow male a couple of yellow females and two black males what is the likely color of the father and

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