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

Molecule synthesis and molecule break-down, Match dehydration reaction and ...

Match dehydration reaction and hydrolysis reaction to molecule synthesis and molecule break-down

Define the principles of periapical surgery (pas), Define the Principles of...

Define the Principles of Periapical Surgery (PAS)   1. Avoid horizontal, sever angled vertical incision Because the collagen fibers of the mucoperiosteum are parallel to the to

Lungs, what is the lungs structure in relation to the excretory system?

what is the lungs structure in relation to the excretory system?

Phenology of different species, Phenology of Different Species The phe...

Phenology of Different Species The phenology of different species present in a community may differ from each other significantly. It is these phenological changes which give

What are colonies and societies, Q. What are colonies and societies? Th...

Q. What are colonies and societies? The Colonies are functional integrated aggregates formed by individuals of the similar species. The Colonies are often confused with a singl

State the aim of neuropsychological assessment, State the aim of Neuropsych...

State the aim of Neuropsychological assessment Neuropsychological assessments aim at specifying in as much detail as possible the functional deficits that exists in a manner th

Determine the long buccal nerve, Long buccal nerve The long buccal nerv...

Long buccal nerve The long buccal nerve is a branch of the mandibular division of the trigeminal nerve which provides sensory innervation to the buccal gingiva and mucosa of th

Metabolism of lipids, The three main forms of fat found in food are glyceri...

The three main forms of fat found in food are glycerides (principally triacylglycerol [triglyceride], the form in which fat is stored for fuel), the phospholipids, and the sterols

How will it impact the rest of the organs, Starting at the esophagus, trace...

Starting at the esophagus, trace the pathway of food through the system. At each organ, indicate anatomical adaptations to the general GI tract tube structure that enhance the spec

What is reverse osmosis, What is reverse osmosis? Give its one application....

What is reverse osmosis? Give its one application. Which two heart sounds are heard through the stethoscope when placed on the chest? When are these sounds produced respectivel

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