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 is ventilation, What is ventilation   A.  An increase in the hydro...

What is ventilation   A.  An increase in the hydrogen ion concentration in the interstitial spaces of the  brain stem leads to an increase in the duration of the respiratory cy

Define the term price mechanism and its working, Define the term price mech...

Define the term price mechanism and how it works. The price mechanism and its working: The above demonstrated diagram assumes the market is into equilibrium, whereby deman

State the term in detail hydrostatic skeleton., State the term in detail hy...

State the term in detail hydrostatic skeleton. Formed from a fluid-filled and closed cavity surrounded by a body wall comprising muscles oriented in different directions. Muscu

Why does bark often break and die naturally, Why does bark often break and ...

Why does bark often break and die naturally? Bark is the mature periderm of the branches, stem and roots. It breaks and dies when these structures grow and thus the peridermal

.development, what is the function of the larval stage in the development o...

what is the function of the larval stage in the development of the young of five named animals

Nerves found in human body, Motor nerves : Nerves which carry impulses fro...

Motor nerves : Nerves which carry impulses from brain or spinal cord to effector organs are called efferent or motor nerves. Stimulation of motor nerves makes the muscles to contr

Define etiology - anorexia nervosa, Define Etiology - Anorexia Nervosa? ...

Define Etiology - Anorexia Nervosa? The exact cause of eating disorders is not known. It is multi factorial in origin in which the personality of the patient, family relationsh

Explain nutritional requirements of microbes, Explain Nutritional Requireme...

Explain Nutritional Requirements of Microbes? Microorganisms require various nutrients and physical factors for their existence and growth. Nutrients are substances used in bio

What is the mendel’s first law, the Mendel's first law? A Mendel's firs...

the Mendel's first law? A Mendel's first law postulates that a characteristic (trait) of an individual is always determined by two factors, one inherited from the mother and th

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