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

Find the magnitude of the acceleration, A proton accelerates from rest in a...

A proton accelerates from rest in a uniform electric field of 650 N/C. At some later time, its speed is 1.3 106 m/s. Find the magnitude of the acceleration of the proton. M/s2

What do you mean by dengue, Q. What is dengue? The Dengue, or dengue fe...

Q. What is dengue? The Dengue, or dengue fever, is an epidemic disease in some countries (for instance, in Brazil), and its most dangerous form is hemorrhagic dengue. It is cau

Essay, “Define nosocomial infections. Discuss the factors that might influe...

“Define nosocomial infections. Discuss the factors that might influence whether a patient may acquire nosocomial infection. How might these risks be reduced?”

How are the main classes of arthropods classified, How are the main classes...

How are the main classes of arthropods classified according to the presence of larval stage in their embryonic development? In crustaceans there are species with direct and oth

Explain factors that alter the speed of enzymatic recations, What are the m...

What are the main factors that alter the speed of enzymatic reactions? The major factors that change the speed of enzymatic reactions are temperature, pH and substrate concentr

Explain alkaline butt -carbohydrate utilization pattern test, Explain Alkal...

Explain Alkaline Butt -carbohydrate utilization pattern test? Alkaline slant (red) and alkaline butt (red) or no change (orange-red) butt- Instead of carbohydrates, peptone in

Sources of variability, Mutations and genetic recombination (in sexually re...

Mutations and genetic recombination (in sexually reproducing organisms) are major sources of variations in natural populations, In this section we will briefly mention the various

Explain about the term- therapeutic, Explain about the term- Therapeutic ...

Explain about the term- Therapeutic Therapeutic approaches that scientists are pursuing include transplanting cells to replace those which are damaged or using growth factors

Protozoa and Fungi compare and contrast, classification schemes and charact...

classification schemes and characterisitics used for this.

Explain about the restriction enzymes, Which of the following is a false st...

Which of the following is a false statement regarding restriction enzymes? A. Restriction enzymes bind to a sequence specific element and disrupt the hydrogen bonds among

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