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

How is the blood typing concerning the abo system, How is the blood typing ...

How is the blood typing concerning the ABO system and the Rh usually done? In the blood typing for the ABO system and the Rh system a blood sample is collected from the person

Inferior alveolar nerve, Inferior alveolar nerve Inferior alveolar ner...

Inferior alveolar nerve Inferior alveolar nerve repositioning to facilitate the placement of endosseous implants posterior to mental foramen is associated with very high incid

What are holandric genes, What are holandric genes? Holandric genes are...

What are holandric genes? Holandric genes are genes situated in the nonhomologous region of the Y chromosome. Holandric genes condition phenotypes that emerge only in men as in

Aba - dormancy, ABA - Dormancy Abscisic acid functions as an antagonis...

ABA - Dormancy Abscisic acid functions as an antagonist of all three promoters like IAA, GAS and Cytokinins. ABA nullifies the activity of GA, IAA and cytokinin right by a)

Demonstration of diversity of plant and animal - taxonomy, Demonstration of...

Demonstration of Diversity of Plant and Animal - Taxonomy A third objective is the demonstration of tremendous diversity of the plant and animal world and its relation to man's

Stratification - aquatic ecosystems, Stratification - Aquatic ecosystems ...

Stratification - Aquatic ecosystems Aquatic ecosystems also exhibit marked stratification. In lake and ocean ecosystems light penetration, temperature and availability of oxyg

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

Signal transduction, What is the initial concentration of Beta-adrenergic r...

What is the initial concentration of Beta-adrenergic receptor in blood?

Examine the cysticerus larva under the microscope, Examine the cysticerus l...

Examine the cysticerus larva under the microscope? Examine the cysticerus larva under the microscope and note the following: i)  Bladder worm consists of a single layered, l

Explain about the saponins, Explain about the Saponins? These are a div...

Explain about the Saponins? These are a diverse group of compounds commonly found in legumes like soybean, lentils, chickpeas, peanuts and alfalfa sprouts. They are also presen

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