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

Infective endocarditis, Infective Endocarditis :  All patients with prosth...

Infective Endocarditis :  All patients with prosthetic valve come under the high-risk category for endocarditis. They need prophylactic antibiotics if any procedure, which is like

Difference between unconditioned & conditioned reflex action, Difference be...

Difference between unconditioned and conditioned reflex action- S.NO .   1. UNCONDITIONED   It is inborn CONDITIONED

Implementation of nursing care for thalassemia patient, Implementation of N...

Implementation of Nursing Care   Prepare the Patient for Diagnostic Evaluation Your role as a nurse is to provide continuing support to the child and family during diagnos

Single pair of alleles, Give three examples of human disorders which are ca...

Give three examples of human disorders which are caused by the action of a single pair of alleles. In every case say whether the harmful allele is dominant or recessive to the non-

Algae, what are the characteristics of algae?

what are the characteristics of algae?

Autotrophic and heterotrophic succession, Autotrophic and Heterotrophic Suc...

Autotrophic and Heterotrophic Succession The succession where initially the green plants are much greater in quantity than the animals, is known as autotrophic succession. Su

What are the grasslands of north america, What are the grasslands of North ...

What are the grasslands of North America and of South America respectively called? The steppe grasslands of North America are known as prairies. The grasslands of South America

Define about the column chromatography, Define about the Column chromatogra...

Define about the Column chromatography? In column chromatography, a tube is filled with the material constituting the stationary phase, plus a solvent (mobile phase). The gener

Precautions should be taken while exercising?, Q. Precautions should be tak...

Q. Precautions should be taken while exercising? Being a diabetic the patient has to take a few precautions. These are: - Initiate the exercise programme gently and then bui

Types of leaves, Types of leaves Collect leaves from such plants as lil...

Types of leaves Collect leaves from such plants as lilies, bamboo, corn, willow and geraniums. View that the monocots (lily, bamboo, corn, sugar cane) have the veins running pa

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