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

Explain the use of benzoates acid, Q. Explain the use of Benzoates acid? ...

Q. Explain the use of Benzoates acid? Benzoic acid, sodium benzoate and the parahydroxy esters (parabens) are used as preservatives. Benzoic acid and its sodium salt, sodium be

Which kind of polarity do fat-soluble substance has, Q. Which kind of polar...

Q. Which kind of polarity do fat-soluble and water-soluble substances respectively have? Ans. Water-soluble substances are behave as polar molecules, i.e., they have e

Define the density of egg, Define the density of egg  The density of e...

Define the density of egg  The density of egg products is not affected by dehydration. When a dried egg product is reconstituted to its natural solids, it has about the same d

Describe cohesion-adhesion theory , Cohesion-adhesion theory describes the...

Cohesion-adhesion theory describes the properties of water which help move it through the plant. Cohesion is the property of water molecules to stick together (held by hydrogen bo

Explain the decolourizing agent - stain technique, Explain the Decolourizin...

Explain the Decolourizing Agent - Stain Technique? 95% ethanol is used as a decolourizing agent. It has two functions - (1) It acts as protein - dehydrating agent, and (2

Explain procambium in primary growing shoot, Explain Procambium in primary ...

Explain Procambium in primary growth in shoot ? The vascular tissue in the shoot is produced from a primary tissue called the procambium, located in the apical meristem. New vas

Describe about water expulsion vesicle, The water expulsion vesicle (also k...

The water expulsion vesicle (also known as a contractile vesicle) is an important osmoregulatory organelle in freshwater protists, which are hyperosmotic to surrounding water. Wate

Preservatives-sulphur dioxide-bisulphites and sulphites, Normal 0 ...

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

Triple fusion, Triple Fusion The fusion process between the other male...

Triple Fusion The fusion process between the other male gamete and the secondary nucleus follows the same pattern as syngamy. In most plants, the polar nuclei are only partial

Spoilage of butter, Q. Spoilage of butter? Butter, which has a high con...

Q. Spoilage of butter? Butter, which has a high content of fat, is subject to rancidity and microbial spoilage due to contaminated cream from which it has been prepared

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