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

Guidelines for diabetic patients for healthy coping, Q. Guidelines for Diab...

Q. Guidelines for Diabetic Patients for Healthy Coping? Generally, a diabetic patient adapts to day to day stress effectively, as they accept their disease condition with the s

Of which substances is chromatin made, Of which substances is chromatin mad...

Of which substances is chromatin made? Chromatin is made of DNA molecules associated to proteins known as histones. Cell Nucleus Review - Image Diversity: chromatin

Types of mental disorder, TYPES OF MENTAL DISORDER - 1 .       Psycho...

TYPES OF MENTAL DISORDER - 1 .       Psychosis 2 .       Neurosis 3.       Personality & character disorders 1 .       PSYCHOSIS - It effects mental capa

Determine the proecss of contrast sense, Contrast sense Sense of contr...

Contrast sense Sense of contrast is the ability to perceive slight changes in luminance between regions which are not separated by definite borders. This is as important as to

Diaphragmatic hernia, Diaphragmatic Hernia: In this condition there  i...

Diaphragmatic Hernia: In this condition there  is a slight herniation ofabdominal organs  (stomach intestine and liver) or extreme protrusion of abdominal contents into the th

What is constitutional unit of proteins, Q. What is constitutional unit of ...

Q. What is constitutional unit of proteins? The constitutional units of proteins are the amino acids.

Explain the periodic table of the elements, Explain the periodic table of t...

Explain the periodic table of the elements ? Periodic Table of the Elements :  An element is a pure substance composed of atoms all of a single kind. It cannot be broken down

Explain the parasite-amoeba, Normal 0 false false false ...

Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4

Define mushroom poisoning, Q. Define Mushroom Poisoning? Mushroom poiso...

Q. Define Mushroom Poisoning? Mushroom poisoning is caused by the consumption of raw or cooked fruiting bodies (mushrooms, toadstools) of a number of species of higher fungl.

Main phases and clinical manifestations of schistosomiasis, Q. What are the...

Q. What are the main phases and clinical manifestations of schistosomiasis? The Schistosomiasis has acute and chronic phases and Days after the infection the cercarial dermatit

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