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

Microscopy, describe the importance,principles,techniques and uses of micro...

describe the importance,principles,techniques and uses of microscopy

Digestive system - buccal cavity, BUCCAL CAVITY - Situated between u...

BUCCAL CAVITY - Situated between upper & lower jaw. It is lined by stratified squamous epithelium. Separated from nasal chamber by palate. Palate forms roof of buccal cav

Blood circulation and respiration, A person is declared to be dead upon the...

A person is declared to be dead upon the irreversible cessation of spontaneous body functions: brain activity, or blood circulation and respiration. However, only about 1% of a per

Describe pulsation in cardian examination, Describe Pulsation in cardian ex...

Describe Pulsation in cardian examination? 1) Apex impulse; lowest and outermost point of definite cardiac pulsation. 2) Prominent pulsations in left parasternal region suggest

Define nutritional requirements for extreme environments, Define Nutritiona...

Define Nutritional Requirements for Extreme Environments? Human beings have been able to survive and work under extreme environments of almost all regions of the earth, from po

How does chewing food help to speed up digestion, How does chewing food hel...

How does chewing food help to speed up digestion? Chewing reduces food to portions small sufficient to be swallowed and enhances the surface area of the food for digestive enzy

What is ammensalim, What is ammensalim? Ammensalism is the ecological i...

What is ammensalim? Ammensalism is the ecological interaction in which an individual harms another without obtaining advanatage. Ammensalism is an inharmonious (negative) ecolo

Explain about the food spoilage, Explain about the Food Spoilage? Foods...

Explain about the Food Spoilage? Foods gradually undergo deterioration or spoilage from the time they are harvested, caught, slaughtered or manufactured. Therefore, delay in th

About ribosome, Which type Ribosome occurs exclusivley in Mitocondria?

Which type Ribosome occurs exclusivley in Mitocondria?

Microtubule assembly, Microtubule assembly: A) generally originates in the ...

Microtubule assembly: A) generally originates in the centrosome. B) occurs only during mitosis. C) occurs randomly throughout the cell. D) is regulated by myosin. E) is inhibited d

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