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

Dna vs protein, Describe the difference between dna and protein. (structura...

Describe the difference between dna and protein. (structural unit, linkage bond, primary and spatial structure)

Find the biosynthesis of acetylcholine, On the overall process of cell-to-c...

On the overall process of cell-to-cell communication within the nervous system, what role does the Ca2+ play in the synapse? A) contributes in the biosynthesis of acetylcholine B)

Birth control - temporary methods, BIRT H CONTROL - Birth control meth...

BIRT H CONTROL - Birth control methods which deliberately prevent fertilization are contraception. These methods may be temporary or permanent. 1 .       TEMPORARY MET

Botany, Sir/Madam, I am a Banquet Farm Owner and face the problem with the ...

Sir/Madam, I am a Banquet Farm Owner and face the problem with the Grass that in November & December the grass does not grow well or disappear beacuse of heavy foot fall. Is their

Explain lipids, Explain lipids? Lipids : Lipids function as energy-sto...

Explain lipids? Lipids : Lipids function as energy-storing molecules such as fats and oils, protective waxes, digestive tract lubricants, heat insulation such as whale blubber

Chi square test and show the chi square test, You are working with plants t...

You are working with plants that have either hairy (dominant) or 'bald' (recessive) flowers. Hairy flowers have little hairs on them that make the flowers look fuzzy, bald flowers

Explain leaf silhouettes, Leaf silhouettes Place a leaf on a sheet of w...

Leaf silhouettes Place a leaf on a sheet of white paper and hold it securely with thumb or finger. Press a piece of natural or artificial sponge next to an ink pad. With short,

Functions of kidney, FUNCTION S OF KIDNEY - 1.      Excretion of nitro...

FUNCTION S OF KIDNEY - 1.      Excretion of nitrogenous wastes. 2.      Maintenance of water balance or osmoregulation. 3.      Acid base balance or regulation of pH.

Explain the term - neurochemical manipulations, Explain the term - Neuroche...

Explain the term - Neurochemical Manipulations Neurochemical and immunological methods have been used to identify groups of neurons in the central nervous system that use speci

Which type of gamete produced by a heterozygous individual, Which is the ty...

Which is the type of gamete produced by a heterozygous individual? What is the genotypical proportion of these gametes? Heterozygous individuals, for instance, AA, produce two

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