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

SYMBIOTIC THEORY, WHAT IS SYMBIOTIC THEORY. PLEASE EXPLAIN THIS TO ME IN A ...

WHAT IS SYMBIOTIC THEORY. PLEASE EXPLAIN THIS TO ME IN A VERY SIMPLE WAY.

Explain phylum-bacteria, Phylum-Bacteria (Eubacteria) Eubacteria,th e t...

Phylum-Bacteria (Eubacteria) Eubacteria,th e true bacteria, are extremely small and they exist in vast numbers virtually everywhere on earth. They range from the upper atmosphe

Predominant feature of steatorrhoea, Q. Predominant feature of steatorrhoea...

Q. Predominant feature of steatorrhoea? A predominant feature is delayed and defective absorption of fat, which results in bulky stools containing large quantities of fat (know

What is vertebrates, What is Vertebrates ? Vertebrates are animals that...

What is Vertebrates ? Vertebrates are animals that have vertebral columns, and include the following classes: Agnatha - jawless fishes (ex. lampreys) Chrondirichthyes - cart

Respiration, comparitive chart on different types of respiration obserbed i...

comparitive chart on different types of respiration obserbed in different type of respiration organ of respiration , types of respiration,energy obtained , by product formed

Assignment, parts involving in circulatory system

parts involving in circulatory system

Fructose does not have an aldehyde group, first of all  bonds of fructose ...

first of all  bonds of fructose was breaked by fehling solution then fructose coverted to aldehydic group then that is react with fehlings solution.

Name two food additives needed to keep food wholesome, a) Name two food ad...

a) Name two food additives needed to keep food wholesome, and say what they do. b) Name two food additives (or types of additive) which are not necessary for keeping fo

Determine the term - techniques visualise anatomy, Determine the term - tec...

Determine the term - techniques visualise anatomy Some techniques visualise anatomy, provide objective confirmation of structural abnormality. These include neuroradiological i

Define hydroxybenzoic acid and hydroxycinnamic acid, Define Hydroxybenzoic ...

Define Hydroxybenzoic acid and Hydroxycinnamic acid? Hydroxybenzoic acid: e.g. ellagic and gallic acids, which are hydrolyzable tannins, present in berries and nuts. Hydroxy

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