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

Cardiac catheterization of mitral stenosis, Q. Cardiac Catheterization of m...

Q. Cardiac Catheterization of mitral stenosis? Cardiac catheterization is rarely needed to diagnose mitral stenosis. An end diastolic gradient more than 5 mm Hg. across mitral

Define potential health benefits from resistant starch, Define Potential He...

Define Potential Health benefits from resistant starch? Like dietary fibre, RS can also play a potential role in helping to maintain or improve health of an individual. As you

Invertebrate zoology, give an example invertebrate phylum/phyla for protopl...

give an example invertebrate phylum/phyla for protoplasmic grade organization

Enzymatic mutation detection method, This technique utilizes an enzyme reso...

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 muta

Nutrition, definition of dietary factors. or what are dietary factors?

definition of dietary factors. or what are dietary factors?

Explain the importance of nicotinic acid, Explain the Importance of nicotin...

Explain the Importance of nicotinic acid Nicotinic acid is successfully used in the therapy of a number of vascular disorders such as vascular spasms, angina pectoris, arthriti

Describe the factors that affect cardiac output, Describe the factors that ...

Describe the factors that affect cardiac output in a female athlete who is speed skating toward the finish line in an Olympic race

Economic dimensions of financing healthcare, Economic Dimensions of Financi...

Economic Dimensions of Financing Healthcare Establishment of adequate healthcare services, accessible/affordable to all sections of the society, is an important function of th

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