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

What are the cells responsible for the antibodies, Q. What are the cells re...

Q. What are the cells responsible for the production of antibodies? The cells that produce antibodies that is the cells of the humoral immune system are the B lymphocytes (B ce

Nursing assessment of acute renal failure, Nursing Assessment If the ...

Nursing Assessment If the cause is due to hypovolemia you may observe the child for dry mucus membrane, poor skin turger, hypotension and oliguria. You should also observe

What are cytokines, What are Cytokines? Cytokines are polypeptides, whi...

What are Cytokines? Cytokines are polypeptides, which influence the proliferation, differentiation, metabolism and activation of cells. There are three areas in cancer that hav

Safety and First-Aid Procedures, How can a family prevent injuries from a f...

How can a family prevent injuries from a fire in the home

Ecology, does jellyfish depend on solarenergy

does jellyfish depend on solarenergy

Animal Respiration, What type of respiration does earthworm has ? In many c...

What type of respiration does earthworm has ? In many complex animals, the cells of internal organs are distant from the outside environment; respiratory systems provide the passag

How are secondary cell walls different from primary cell, How are secondary...

How are secondary cell walls different from primary cell walls? Primary cell walls are assembled on the surface of the plasma membrane whereas the cell is growing. They can gro

What are some things that have algae in them, What are some things that hav...

What are some things that have algae in them? Yeast is considered Algae.

What is choanocytes, Which one of the following statements about all the fo...

Which one of the following statements about all the four of Spongilla, Leech, Dolphin and Penguin is correct? 1. Penguin is homiothermic while the remaining three are poikilothe

Endothelium derived relaxing factors, Vasoactive substances such as Endothe...

Vasoactive substances such as Endothelium Derived Relaxing Factors (EDRF). Prostacyclin and Endothelin can be formed in the vascular endothelium. Endothelial dysfunction can lead t

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