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

Alcohol related emergencies, Alcohol Related Emergencies: Alcohol is o...

Alcohol Related Emergencies: Alcohol is one of  the most serious health problems. Alcoholism is defined as a primary or chronic disease with genetic, psychological and environ

What is eukaryotic cells , What is Eukaryotic Cells ? Eukaryotic Cel...

What is Eukaryotic Cells ? Eukaryotic Cells :  Most cells of higher organisms, and many unicells, contain a nucleus. Organisms whose cells contain a nucleus are called euka

Which cells produced in first stage of embryonic development, Q. What are t...

Q. What are the cells produced in the first stage of the embryonic development called? The cells that result from the cleavage (the first stage of the embryonic development) ar

Common disease of goats, COMMON DISEASE OF GOATS - 1 .       ANTHRAX ...

COMMON DISEASE OF GOATS - 1 .       ANTHRAX 2 .       FOOT & MOUTH DISEASE 3 .       CONTAGIOUS PNEUMONIA - Characterized by pneumonia & pleurisy, cough, snee

Use of protective eyewear as personal protective equipment, Q. Use of Prote...

Q. Use of Protective Eyewear as personal protective equipment? Protective lenses must be worn by dental personnel: 1. when performing procedures that can cause spatter or ae

Explain trapezoidal full mucoperiosteal flaps, Explain Trapezoidal Full Muc...

Explain Trapezoidal Full Mucoperiosteal Flaps a) Similar to rectangular flap except: The 2 vertical incisions are at obtuse angle with the Horizontal incision, b) Mostly

What are the five kingdoms, Q. What are the five kingdoms into which living...

Q. What are the five kingdoms into which living beings are divided? And which group of living being is out of this classification? The five kingdoms of living beings are the ki

Polychaete, Polychaete, Mollusca and Echinodermata Polychaete and Mol...

Polychaete, Mollusca and Echinodermata Polychaete and Mollusca The polychaetes have a trochophore larva. A common trochophore larva is top shaped bearing a tuft of cilia

Determine the chain in the dna molecule, Which type of chemical bond mainta...

Which type of chemical bond maintains the pairing of each chain in the DNA molecule? To produce the DNA molecule, purine bases bind to pyrimidine bases by intermolecular bonds

Prevention and control of shigella food borne infection, Prevention and Con...

Prevention and Control of Shigella food borne infection The control of  Shigella food borne infection is similar to that of salmonellae; avoiding contamination of foods by ani

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