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

Tolerance range, Maximum and minimum range of an animal with the factors of...

Maximum and minimum range of an animal with the factors of Ph Salinity Pressure Relative human

Which substances transferre from the mother to the fetus, Q. What are the m...

Q. What are the major substances transferred from the mother to the fetus through the placenta? And from the fetus to the mother? From the mother to the fetus the major transfe

Blood groups, BLOO D GROUPS- In R.B.C. on plasmalemma antigen or agglu...

BLOO D GROUPS- In R.B.C. on plasmalemma antigen or agglutinogen present form antibodies or agglutinines. Blood groups are based on antigens. 1 .      ABO SYSTEM -

Explain pennate diatomsrole in diatoms, Explain Pennate diatomsrole in diat...

Explain Pennate diatomsrole in diatoms? Pennate diatoms have bilateral symmetry, which presents side-by-side mirror images if divided down the middle or centerline, as seen

Change in structure of a protein, Q. Is it expected that a change in the pr...

Q. Is it expected that a change in the primary, in the secondary or in the tertiary structure of a protein will produce furthermore functional consequences? Any change of the p

Canned and retort pouched meat products, C a nned and Retort Pouched Meat...

C a nned and Retort Pouched Meat Products Canning as a process contributes to preservation, storage and distribution of food supplies. Production of thermally processed meat

Precautions by dangers of working in labs with animals, The dangers of work...

The dangers of working in labs with animals will be minimised if the following precautions are taken: 1. Wash all dissection instruments in disinfectant after use. 2. Never

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