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

Mammalian lungs, Definition,functions and features of a mammalian lungs

Definition,functions and features of a mammalian lungs

Ablation experiment, Ablation experiment  is an experiment designed to prod...

Ablation experiment  is an experiment designed to produce an animal deficient in one or the few cell types, in order to study cell lineage or the cell function. The logic is to mak

What is xaxim, What is xaxim? Most pteridophytes have subterraneous ste...

What is xaxim? Most pteridophytes have subterraneous stems similar to the substrate called as rhizomes. Xaxim is a kind of pteridophyte with an aerial stem in generally perpend

Explain the peripheral resistance, Explain the Peripheral Resistance Re...

Explain the Peripheral Resistance Resistance offered by arterioles or resistance vessels, as you have read above, is termed as peripheral resistance. Changes in peripheral resi

What is a photosynthesis, Photosynthesis is a process used by plants and ot...

Photosynthesis is a process used by plants and other organisms to change the light energy captured from the sun into chemical energy which can be beneficial to fuel the organism's

Explain functional properties of protein hydrolysis, Explain functional pro...

Explain functional properties of protein hydrolysis Hydrolysis of food proteins using proteases (trypsin, chymootrypsin, papain and thermolysin) alters their functional propert

What is cryopreservation, Name the location and function of Meibomian gland...

Name the location and function of Meibomian glands in the human eye. a) What is cryopreservation? Give its single use. b) Commercial importance of cryopreservation is relate

Define atherosclerosis, Q. Explain Atherosclerosis? Atherosclerosis is ...

Q. Explain Atherosclerosis? Atherosclerosis is one of the most important causes of morbidity and mortality in developed as well as in developing countries. Atherogenesis occurs

Dysrhythmias - complications of cardiac surgery, Dysrhythmias Usually ...

Dysrhythmias Usually occur during the initial 24-72 hours, but may occur later on also. The causes may be ventricular irritability due to manipulations of heart durin

What is the histological nature of the glands, What is the histological nat...

What is the histological nature of the glands? How are they formed? The glands are epithelial tissues. They are made of epithelium that throughout the embryonic development inv

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