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

Name the three main arthropod classes, How are the three main arthropod cla...

How are the three main arthropod classes characterized according to the body division? In crustaceans and arachnids the head is fused with the thorax forming the cephalothorax.

Explain requirement for simple staining of bacterial culture, Explain Requi...

Explain Requirements for Simple Staining of Bacterial Cultures Culture: 24 hours bacterial culture of E. coli and Bacillus on nutrient agar or in broth. Reagents: Methylene

Genetic or acquired deficiency of the vitamin k metabolism, Q. Why are some...

Q. Why are some kinds of hemorrhagic diseases caused by genetic or acquired deficiency of the vitamin K metabolism? Deficiency of vitamin K predisposes to hemorrhages since thi

What are the physical propertie of sol, What are the physical propertie of ...

What are the physical propertie of sol Sols resemble liquids in their main physical properties - that is, they flow and they do not show rigidity of form. However, when a sol a

Explain the working of senses: touch, Explain The working of Senses: Touch,...

Explain The working of Senses: Touch, Hearing, Equilibrium ? The senses are detected by sense organs, collections of receptor cells, or cells closely associated with them. A m

Explain oleic - linoleic fats, Oleic - Linoleic Fats Fats in this grou...

Oleic - Linoleic Fats Fats in this group are the most abundant. The oils are all of vegetable origin and contain large amounts of oleic and linoleic acids, and less than 20%

Explain feeding schedule in infancy, Explain Feeding Schedule in Infancy? ...

Explain Feeding Schedule in Infancy? 1) Milk feed 5-6 times/day. Continue breast-feeding. Exclusive breast-feeding up to 6 months even during illness. Follow demand feeding.

Report writing, tell me how to write a report in molecular biology?

tell me how to write a report in molecular biology?

Sequencing of dna fragments, Q. If a fragment of nucleic acid has a nucleot...

Q. If a fragment of nucleic acid has a nucleotide sequence TAC can one assert that it is an anticodon or a codon? A nucleic acid having a TAC sequence surely is not tRNA it is

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