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

Excretion in protozoans, Excretion in Protozoans Excretion of metaboli...

Excretion in Protozoans Excretion of metabolic wastes is dope almost exclusively by diffusion. All protozoans are ammonotelic i.e., the end product of their nitrogen metabolis

Vitamins and minerals requirement in chronic diarrhoea, Q. Vitamins and min...

Q. Vitamins and minerals requirement in chronic diarrhoea? Loss of vitamins is related to the degree of mucosal damage in chronic diarrhoea, which in turn impairs absorption an

Investigating, What is scientific investigation?

What is scientific investigation?

Explain the parasite-amoeba, Normal 0 false false false ...

Normal 0 false false false EN-US X-NONE X-NONE MicrosoftInternetExplorer4

Cleavage - development biology, Cleavage - Development Biology Cleavag...

Cleavage - Development Biology Cleavage or segmentation is a series of cell divisions of the fertilized .egg through which it is converted into a multicellular structure, call

What are dopaminergic neurons, Q. What are Dopaminergic neurons? Dopam...

Q. What are Dopaminergic neurons? Dopamine Dopaminergic neurons occur in two closely connected groups: Ventral tegmental area (VTA) of the midbrain and Substantia nigra,(i

Show symptoms and signs of hyperthyroidism, Q. What are some symptoms and s...

Q. What are some symptoms and signs found in patients with hyperthyroidism? The hormones made by the thyroid gland stimulate the basal metabolism of the body in hyperthyroidism

Bacteria and archea, Based on the scientific name. Streptococcus agalactia...

Based on the scientific name. Streptococcus agalactiae what morphology would you expect these cells to have?

Classification of agro-industrial byproducts, Classification of agro-indust...

Classification of agro-industrial byproducts Based on their nutrient content agro-industrial byproducts can be divided into: 1.  Feed low in fiber and low in protein. These ar

What are the major cells of which poriferans are made, Q What are the major...

Q What are the major cells of which poriferans are made? Sponges have their external wall covered by flat cells called pinacocytes and having pores well-delimited by special ce

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