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 to increased co2 levels in the atmosphere, Increased CO 2 levels in th...

Increased CO 2 levels in the atmosphere are correlated with rising global temperatures, leading many scientists to believe that the first phenomenon has caused the second. What wo

Factors associated with accident, Factors Associated With Accident Co...

Factors Associated With Accident Cost in Administrative Functions Loss of prestige. Costs for notification, investigation and legal action. Loss of pu

Explain the occurrence of vitamin B6, Explain the Occurrence of vitamin B 6...

Explain the Occurrence of vitamin B 6 Vitamin B 6 activity is attributed to the 3 compounds-pyridoxol (pyridoxine), pyridoxal and pyridoxamine, generally comprised in the vit

Explain about the diabetes mellitus, Explain about the Diabetes mellitus? ...

Explain about the Diabetes mellitus? You are aware that diabetes may be diagnosed as an exaggerated response in blood glucose concentration following ingestion of a fixed amou

Show the use of incineration for sterilization, Q. Show the Use of Incinera...

Q. Show the Use of Incineration for sterilization? This is an excellent method for rapidly destroying materials such as soiled dressings, animal carcasses, bedding and patholog

How can the hypothesis that asserts that chloroplasts, How can the hypothes...

How can the hypothesis that asserts that chloroplasts as well as mitochondria were primitive prokaryotes that associated in mutualism with primitive anaerobic eukaryotic cells be c

Define drug effects on carbohydrate metabolism, Define drug effects on Carb...

Define drug effects on Carbohydrate metabolism? Carbohydrate metabolism: Hypoglycemic drugs such as insulin and sulphonylureas are prescribed because of their ability to increa

Why theories are import, Theories are import because they can be used to a....

Theories are import because they can be used to a. Test research hypothesis b. Develop questions which will facilitate more effective research c. Conduct more meaningful and useful

Amphibians, respiratory system in amphibains

respiratory system in amphibains

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