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

Define some usual doubts of children related to food, Define some Usual Dou...

Define some Usual Doubts of Children related to Food? Before embarking yourself to go through this unit, here is an activity for you to perform, Talk to 2-3 school' boys and gi

Discuss the biological concepts of the migration, The bar-tailed godwit, Li...

The bar-tailed godwit, Limosa lapponica baueri, or kuaka, is a wading shorebird seen on marshy estuaries and wetlands during the New Zealand summer. Populations of this species emb

Explain database search, Database Search: Once an open reading frame or th...

Database Search: Once an open reading frame or the partial amino acid sequence has been pre-determined, the investigator compares sequence with others in the databases using a com

Define working of body surface in excretion of zinc in human, Define workin...

Define working of Body surface in excretion of zinc in human? Loss of zinc occurs due to the exfoliation of skin and sweating (0.7 -1.0 mg/day). Another route of zinc loss is h

The warmest parts of the body?, What are likely to be (a) the coldest, (b) ...

What are likely to be (a) the coldest, (b) the warmest parts of the body? The extremities of the body (hands and fingers, feet and toes, ears and nose) are likely to be the col

Sulphur dioxide emissions, State two ways in which sulphur dioxide emission...

State two ways in which sulphur dioxide emissions from coal-fired generating stations could be reduced. Sulphur dioxide emissions can be reduced by fitting desulphurization pl

Spiracular control - respiration, Spiracular Control - Respiration ...

Spiracular Control - Respiration Normal 0 false false false EN-IN X-NONE X-NONE MicrosoftInternetExplorer4

Microscopes, differentiate between scanning electron microscope and a trans...

differentiate between scanning electron microscope and a transmission electron microscope in terms of their functions

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