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

Carbohydrates requirements for ulcerative colitis, Q. Carbohydrates require...

Q. Carbohydrates requirements for ulcerative colitis? Carbohydrates: They form the easily absorbable source of energy. Bulk-producing vegetables are restricted so as to allow b

Pedigree charts, How do I complete a pedigree chart for genetics?

How do I complete a pedigree chart for genetics?

What is the function of the citric acid cycle, What is the function of the ...

What is the function of the citric acid cycle? The hnction ofthe  citric acid cycle can be discussed as  follows. The  intermediates of citric acid cycle are used as  precursor

Body fluids of marine teleost, Body Fluids of Marine Teleost The body ...

Body Fluids of Marine Teleost The body fluids of marine teleost are hypotonic to seawater, so these fishes lose water to the environment especially across the gill epithelium.

Define factors affecting absorption of dietary iron, Define Factors affecti...

Define Factors affecting Absorption of Dietary Iron? There are mainly four factors that determine iron bioavailability absorption from the diet. These include: i) Form of ir

Define epithelial tissue, Epithelial tissue the cells in animals which are...

Epithelial tissue the cells in animals which are closely packed in either single or multiple layers, and which cover internal and external surfaces both of the animal body. It is

Define reduction of blood lipids, Define Reduction of blood lipids? Thi...

Define Reduction of blood lipids? This effect have been especially observed with saponins. Diets containing saponin-rich foods (300-500 mg saponins/day) e.g. soya, alfalfa, chi

Explain wind pollinated flowers, Wind pollinated flowers are: 1. Small, ...

Wind pollinated flowers are: 1. Small, brightly coloured, producing large number of pollen grains 2. Small, producing large number of dry pollen grains 3. Large producing

J-shaped growth curve, J-shape d - It shows 2 phases - (i) Lay ph...

J-shape d - It shows 2 phases - (i) Lay phase (ii) Exponential phase

Cycling of nutrients, Pick up just a tiny bit of soil and you will discover...

Pick up just a tiny bit of soil and you will discover a whole new world. Researchers have found that one pinch of soil may contain more than 30,000 protozoa, 50,000 algae, 400,000

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