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

Glycolysis, Glycolysis Oxidation of glucose is called as glycolysis.Glu...

Glycolysis Oxidation of glucose is called as glycolysis.Glucose is oxidized to either lactate or pyruvate. Under aerobic conditions and the dominant product within most tissues

Infants with maple syrup urine disease, Q. Infants with maple syrup urine d...

Q. Infants with maple syrup urine disease? Infants with MSUD appear normal at birth. Once they are fed on protein containing feed, they start showing the classic symptoms. Neur

Bronchial asthma, Bronchial Asthma Bronchial Asthma is characterized ...

Bronchial Asthma Bronchial Asthma is characterized by increased responsiveness of the trachea and bronchi to various stimuli that cause narrowing of the airways and difficu

What is protein synthesis, What is Protein Synthesis? Protein Synthesi...

What is Protein Synthesis? Protein Synthesis :  Protein synthesis begins in the nucleus, with the formation of mRNA from a DNA template through transcription. The function of

Explain about the canning of food, Explain about the Canning of Food? Y...

Explain about the Canning of Food? You must have seen the markets flooded with canned products. Can you name a few of such products? Yes the canned juices, canned aerated drink

Polarity in regeneration, Polarity in Regeneration A distinct polarit...

Polarity in Regeneration A distinct polarity or gradient exists in planarians (and in other animals). The meaning of this is that in any piece or segment of the body; its ant

Solar energy, It is an inexhaustible and population free source of energy. ...

It is an inexhaustible and population free source of energy. This energy maintains temperature of the earth. It causes current in the atmosphere and in ocean. The total solar energ

Define about the resistant starch, Define about the Resistant Starch? U...

Define about the Resistant Starch? Until 1980's starch was thought to be completely hydrolyzed and absorbed from the small intestine of man, independent of its source, type and

Plasmid, Plasmid  is the class of the circular, extrachromosomal, autonomou...

Plasmid  is the class of the circular, extrachromosomal, autonomously replicating, DNA elements found in number of bacteria. Contain origins of the replication to ensure their main

Explain pasteurization, Explain Pasteurization The act or process of h...

Explain Pasteurization The act or process of heating a beverage or other food, to a specific temperature for a specific period of time in order to kill microorganisms that cou

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