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

Constrictive pericarditis-pericardial diseases, Constrictive Pericarditis :...

Constrictive Pericarditis :  It is usually the end stage of inflammatory process involving pericardium. In developing world, infection with mycobacterium tuberculosis is still com

Explain about the food product development, Explain about the Food product ...

Explain about the Food product development? Food product development is often commodity related. This type of research needs to be carried out in the pilot plants with the equi

Describe about the tertiary prevention - food allergy, Describe about the T...

Describe about the Tertiary Prevention - food allergy? The tertiary allergy prevention, as you already know, focus on patients with symptoms. Hence, the strategies will target

Explain culture characteristics of molds, Q. Explain Culture Characteristic...

Q. Explain Culture Characteristics of Molds? The gross appearance of a mold growing on a food often is sufficient to indicate its class or order. Some molds are loose and fluff

How neural impulse is transmitted from one cell to another, Q. What is the ...

Q. What is the structure through which the neural impulse is transmitted from one cell to another? What are its parts? The structure through which the neural impulse passes fro

Rationale based on theories of miasma or contagion, Early campaigns of the ...

Early campaigns of the 19th century that focused on sanitation, hygiene, housing, and nutrition had little effect in controlling communicable disease due to flawed rationale based

Growth rate in animals, GROWTH RATE IN ANIMALS - Growth rate is differe...

GROWTH RATE IN ANIMALS - Growth rate is different at different periods of life. Growth period in human may be studied in 5 steps - (i) Prenatal - In gestation period fo

Describe bentham and hookers system, Q. Describe Bentham and Hookers System...

Q. Describe Bentham and Hookers System? Bernard de Jussieu (1699-1776) tried to classify the plants in Royal Garden, Paris. During this exercise he developed a system of classi

What do you mean by oesophagitis, Q. What do you mean by Oesophagitis? ...

Q. What do you mean by Oesophagitis? We already know that oesophagus is a muscular tube 25 cm in length and basically helps in transporting the food from the mouth to stomach,

Biological sickness of soil, what is biological sickness of soil and its ma...

what is biological sickness of soil and its management

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