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

In hormonal terms why does menses occur, In hormonal terms why does menses ...

In hormonal terms why does menses occur? Menses is the endometrial monthly desquamation that happens as the estrogen and progesterone levels fall after the regression of the co

Interspecific sterility, In interspecific sterility, the failure in mating ...

In interspecific sterility, the failure in mating occurs because of inability of the sperm to reach the egg in animals and the pollen to reach ovules in plants. In plailts interspe

What is the ecological role of earthworms, Q. What is the ecological role o...

Q. What is the ecological role of earthworms? Earthworms have an important ecological role as they eat decomposing organic material. They also dig tunnels in the subsoil allowi

What are the symptom of short bowel syndrome, Q. What are the Symptom of sh...

Q. What are the Symptom of short bowel syndrome? Diarrhoea is the main symptom of short bowel syndrome. Other symptoms include: • Anorexia • Steatorrhoea • Heart bum a

Vaccines for gas infections, In developing countries because of poor compli...

In developing countries because of poor compliance, poverty, ignorance, primary and secondary prevention strategies are difficult to implement properly. Also in western countries (

Determine the two different alleles, What is meant by saying that in relati...

What is meant by saying that in relation to a given trait conditioned by a gene with two different alleles the gametes are always "pure"? To say that gametes are pure means tha

Insects - hormones in growth and reproduction, Insects - Hormones in Growth...

Insects - Hormones in Growth and Reproduction In insects hormones regulate moulting and metamorphosis. The larvae or nymphs which hatch out of the eggs undergo regular moultin

Can you explain diabetes mellitus, Q. What is diabetes mellitus? Diabet...

Q. What is diabetes mellitus? Diabetes mellitus is the disease caused by action or deficient production of insulin and the consequent high blood glucose level and low glucose u

Atypical distribution of pleural fluid, Q. Atypical Distribution of Pleural...

Q. Atypical Distribution of Pleural Fluid? i) Lamellar effusion: These are shallow collections of fluid between the chest wall and the lung surface. ii) Subpulmonic

Trans myocardial laser re vascularisation, Trans Myocardial laser re vascul...

Trans Myocardial laser re vascularisation (TMLR) :  Patients who have severe angina and diffuse coronary artery disease with no graftable vessels are at limes subjected to TMLR. I

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