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

Describe class echinoidea in animal kingdom, Describe Class Echinoidea in a...

Describe Class Echinoidea in animal kingdom? Sand dollars and sea urchins either have flattened disc-like bodies, or are somewhat spherical. While they lack the typical five-ar

Define the skeletal muscle, Define the Skeletal muscle A. Binding of ca...

Define the Skeletal muscle A. Binding of calcium ion to its receptor site on the actin molecule blocks the attachment of the head of the tropomyosin molecule to its binding sit

Explain adverse effects of zidovudine, Explain Adverse effects of Zidovudin...

Explain Adverse effects of Zidovudine  It include anemia, neutropenia, nausea, vomiting, headache, fatigue, confusion, malaise, myopathy, hepatitis, and hyperpigmentation of or

Define bioavailability of pyridoxine, Define Bioavailability of pyridoxine?...

Define Bioavailability of pyridoxine? A recent review by Gregory confirms that bioavailability of vitamin B 6 in a mixed  diet is about 7570, with approximately 8% of this tot

What is interpretation of stress echocardiography, Q. What is Interpretatio...

Q. What is Interpretation of Stress Echocardiography? The standard approach to interpretation is qualitative. The attendant need for training and problems posed for reproducibi

What are compound eyes, Q. What are compound eyes? Arthropods have comp...

Q. What are compound eyes? Arthropods have compound eyes made of several visual units called as ommatidia. Each ommatidium transmits visual information through the optic nerve

Who is aristotle - ancient greeks and romans, Who is Aristotle - the Stagir...

Who is Aristotle - the Stagirite? The ptodigious activity of Aristotle (384-323 B.C.) marks the climax of the Golden Age of Greece. The very existence of his works proves not s

How yersinia enterolytica infection occurs, Normal 0 false fa...

Normal 0 false false false EN-IN X-NONE X-NONE MicrosoftInternetExplorer4 How Yersinia enterol

Amoeboid protozoan, Amoeboid Protozoan Amoeboid protozoan are distingu...

Amoeboid Protozoan Amoeboid protozoan are distinguished by the presence of flowing extensions of their body known as pseudopodia. These are used for feeding and locomotion. Th

Sterigmatocystin, These mycotoxins are structurally and biologically relat...

These mycotoxins are structurally and biologically related to the aflatoxins, and like the latter, they cause hepatocarcinogenic activity in animals. Among the organisms that prod

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