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

CORAL REEF., A COMPLETE SEMINAR TALK FOR CORAL REEF

A COMPLETE SEMINAR TALK FOR CORAL REEF

Define the food system in space, Define the Food System in space? Mercu...

Define the Food System in space? Mercury (1961-1963) astronauts had to eat bite-sized cubes, freeze dried powders, and semi liquids stuffed in aluminium tubes. For most asbonau

Cell, how does a cell work ?

how does a cell work ?

Adrenocortical steroids - vertebrates, Normal 0 false false ...

Normal 0 false false false EN-IN X-NONE X-NONE MicrosoftInternetExplorer4

Electron transport , 1) It is the series of coupled oxidation/reduction rea...

1) It is the series of coupled oxidation/reduction reactions where the electrons are passed like hot potatoes from one membrane-bound protein/enzyme to another before being finally

Explain the oligosaccharides (dp: 3-9) - carbohydrates, Explain the Oligosa...

Explain the Oligosaccharides (DP: 3-9) - carbohydrates? Oligosaccharides consists of short chains of monosaccharide units joined by covalent bonds. The glycosidic bond may be α

Female reproductive system from the external vulva, Q. What are the anatomi...

Q. What are the anatomical relationships between the organs of the female reproductive system from the external vulva to the ovaries? The external female genitalia are called t

Number of chromosomes, NUMBER Benden and Boveri first indicated tha...

NUMBER Benden and Boveri first indicated that number of Chromosomes is definite in each organism. (1) Haploid (n) - One set of Chromosomes. i.e. one Chromosome of each

Bacterial diseases-brucellosis, Brucellosis Brucellosis among cattle, ...

Brucellosis Brucellosis among cattle, buffaloes, sheep, goats, pigs produces heavy economic losses due to abortions in late pregnancy, infertility and reduction in productivit

Explain about the whey protein concentrates, Explain about the Whey Protein...

Explain about the Whey Protein Concentrates? You already know that whey is the residual liquid substance that is obtained by separating the coagulum from milk during cheesemaki

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