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

Explain cori cycle , Under the limiting oxygen conditions experienced in ...

Under the limiting oxygen conditions experienced in by vigorous exercise, the configuration of NADH by glycolysis exceeds the ability of the respiratory chain to oxidize it back to

Types of neurons, TYPE S OF NEURONS - The neurons are classified on th...

TYPE S OF NEURONS - The neurons are classified on the basis of their structure and function. (A) On the basis of the structure, the neurons are of four types.

Explain about the nutritional requirements, Explain about the Nutritional R...

Explain about the Nutritional Requirements? Nutritional requirements are defined as 'intake levels of nutrients that meet specified criteria of adequacy such as normal growth,

Advantages and disadvantages of tissue valves, Advantages of Tissue Valv ...

Advantages of Tissue Valv es   Need for anticoagulation therapy is only for a short duration in xenograft and only there is rare need for anticoagulation therapy in au

The chance of fertilisation in humans, Explain why the chance of fertilisat...

Explain why the chance of fertilisation in humans is restricted to only a few days each month. Sperms can fertilise an ovum for up to about three days after entering the female

What is a membrane, What is a membrane? Membrane is any delicate sheet ...

What is a membrane? Membrane is any delicate sheet that divides one region from another blocking or permitting (selectively or completely) the passage of substances. The skin,

What is the hematopoiesis, Q. What is the hematopoiesis? Hematopoiesis ...

Q. What is the hematopoiesis? Hematopoiesis is the formation of blood cells with other constituent elements of the blood. Q. Where does hematopoiesis occur? Hematopoies

Cardiovascular endurance component for physical fitness, Define Cardiovascu...

Define Cardiovascular endurance Component for physical fitness? In our previous section on work capacity, concepts of aerobic capacity Vo2 max were described. Cardiovascular en

What is translocation, What is Translocation? Sometimes pieces of two c...

What is Translocation? Sometimes pieces of two chromosomes that are not homologous break off at the same time. If the broken pieces are re-attached to the opposite (in this cas

Why water and electrolytes are important for exercise, Why Water and Electr...

Why Water and Electrolytes are important for exercise? Water and electrolytes are important for exercise performance as they maintain blood volume and osmolality, thermoregulat

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