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

Disorder of parathyroid function, Disorder of Parathyroid Function: Th...

Disorder of Parathyroid Function: The disorders of parathyroid function may  either  lead  to increased production or decreased production of parathormone.  Hypoparathyroid

Describe the process of coronary atherosclerosis, Q. Describe the process o...

Q. Describe the process of coronary atherosclerosis? Atherosclerosis is an intimal disease of arteries ranging in size from the aorta to the epicardial coronary artery i.e. cha

What are gonads, What are gonads? What are the male and the female gonads i...

What are gonads? What are the male and the female gonads in humans? Gonads are the organs that make gametes. They have the germ cells that undergo division and generate gametes

What is echocardiography, Q. What is Echocardiography? Ans. Echoca...

Q. What is Echocardiography? Ans. Echocardiography has become an established and powerful tool for diagnosing presence of CAD and defining its consequence. It can help in

What is the need of monitoring and surveillance, What is the need of Monito...

What is the need of Monitoring and Surveillance? Monitoring, we know, is the act of observing something and sometimes keeping a .record of it while surveillance is a repeated s

Disorders related to nutrition and digestive system, DISORDE R RELATED TO ...

DISORDE R RELATED TO NUTRITION AND DIGESTIVE SYSTEM Malnutrition Malnutrition is improper intake of food. Malnutrion covers both undernutrition and overnutrition. Und

Define precaution-estimation of nitrogen and protein content, Define Precau...

Define Precautions - Estimation of Nitrogen and Protein Content? 1. The apparatus should be airtight to prevent leakage of NH 3 . 2. The tip of the delivery tube should be i

Explain the heart rate monitoring (hrm) method, Explain the Heart Rate Moni...

Explain the Heart Rate Monitoring (HRM) Method? HRM is a method to measure the daily energy expenditure of free-living individuals, based on the relationship of heart rate and

Experiment turn out differently then you expected, If the results of an exp...

If the results of an experiment turn out differently then you expected, what does that mean? A. The experiment was a failure B. You did not follow scientific method C. You should e

What benefits can commensalism offer to a species, What benefits can commen...

What benefits can commensalism offer to a species? Commensalism may include obtainment of food (for instance, the innocuous bacteria of the human gut), shelter or support (epip

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