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 any three adaptations of mangroves, Name and describe any three ad...

Name and describe any three adaptations of mangroves to the conditions prevailing in the Sunderbans.

Anatomy of the brain, In order to be able to study the neural basis of perc...

In order to be able to study the neural basis of perceptual and cognitive functions, it is necessary to have some knowledge of brain anatomy (i.e., what the names of different brai

How are living beings divided into two groups?, Q According to the cellular...

Q According to the cellular organization how are living beings divided into two groups? Cellular beings are divided into two groups, unicellular beings and the prokaryotes whos

Explain the periodontal consideration, Explain the Periodontal consideratio...

Explain the Periodontal consideration Periodontal consideration would include: Hard Calculus Build up:   Any index assessing the amount of calculus build up can be used .

Ecology and ecosystem, Ecology and Ecosystem The concepts of environme...

Ecology and Ecosystem The concepts of environment, ecology, ecosystem, energy flow and nutrient cycling in the foundation course on Science and Technology. As you are aware ec

Explain about the holmgren''s wools, Explain about the Holmgren's  Wools  ...

Explain about the Holmgren's  Wools   This is a matching test  of  coloured pieces of  wool. While this  is not  a highly  favored method  of  testing  it does  show  serious d

Explain about the fructnns as food ingredients, Explain about the Fructnns ...

Explain about the Fructnns as Food Ingredients? Oligosaccharides have several functional/technological attributes that give them a potential for incorporation into foods. They

Explain the term- horizons, Explain the term- Horizons These processes ...

Explain the term- Horizons These processes take a long time- may be a few thousand years to create a soil. As the time passes, the soil matures and generally becomes deeper and

Which statement is false, Which is FALSE? Diastereomers: Select one: ...

Which is FALSE? Diastereomers: Select one: a. Have the same molecular weight b. Have the same connectivity c. Are mirror images d. All of the above e. None of the

What are vitamins, Q. What are vitamins? What are the major vitamins needed...

Q. What are vitamins? What are the major vitamins needed by humans? Most vitamins are coenzymes fundamental substances for the enzyme functioning that are not produced by the o

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