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

Describe the complete transposition of great arteries, Describe the Complet...

Describe the Complete transposition of great arteries ? Lethal, relatively frequent malformation. Without treatment, it results in 30 per cent mortality within the first week o

Bioclimatology effect of climate on distribution of organism, Bioclimatolog...

Bioclimatology: This is the study of effect of climate on distribution of organisms. Bioclimatology is the inter-disciplinary field of science which studies the interactions betwe

Chronic bronchitis, Chronic Bronchitis Chronic Bronchitis is defined ...

Chronic Bronchitis Chronic Bronchitis is defined clinically as hypersecretion of mucus and recurrent episode of productive cough for a period of 3 months per year at least tw

Aschelminthes, in what part of the human body aschelminthes found?

in what part of the human body aschelminthes found?

Guidelines for the family members for healthy coping, Q. Guidelines for the...

Q. Guidelines for the Family Members for Healthy Coping? - Patient and the family members should discuss the situation, change in role,time distribution, share of money and oth

Explain why goblet cells are non-functional, If for some reason our goblet ...

If for some reason our goblet cells are non-functional, this will adversely affect: 1. Production of somatostatin 2. Secretion of sebum from the sebaceous glands 3. Matura

Nemerteans - regeneration in invertebrates, Nemerteans - Regeneration in In...

Nemerteans - Regeneration in Invertebrates Nemerteans as well possess remarkable regenerative ability, as even a small fragment can regenerate into a complete worm. Nemerteans

Type of disease identified chromosomes present in karyotype, Which type of ...

Which type of genetic disease can be identified from the visual analysis of the number of chromosomes present in a karyotype? The counting and the identification of chromosomes

Ephemeral fever, Ephemeral fever It is also known as three days sicknes...

Ephemeral fever It is also known as three days sickness and affected animals suffer from pyrexia, muscular stiffness and lameness. Et i o l o g y :

Which kind of polarity do water-soluble have, Which kind of polarity do wat...

Which kind of polarity do water-soluble and fat-soluble substances respectively have? Water-soluble substances are polar molecules, i.e., they have electrically charged areas.

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