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

Is osmolarity of the interstitial fluid increase or decrease, If the osmola...

If the osmolarity of plasma (inside a capillary) is 300 mOsM, the osmolarity of the tissue fluid (interstial fluid) is 400, and the membranes are permeable to water but not to salt

What is the thermopenetration concept, What is the thermopenetration concep...

What is the thermopenetration concept? The pasteurization of products packed in glass containers leads to a problem of a specific nature, which is referred to as 'thermopenetra

How is the respiratory system in aquatic molluscs, How is the respiratory s...

How is the respiratory system in aquatic molluscs divided? What adaptive respiratory structure do terrestrial molluscs present? Aquatic molluscs oxygenate their blood through g

Water supply, A rural town in an established agricultural area with no pros...

A rural town in an established agricultural area with no prospects for extensive development is located along a river that is used as the source of water supply for the community.

Human diseases caused by virus, Q. What are some human diseases caused by v...

Q. What are some human diseases caused by virus and what are their respective modes of transmission? The major viral diseases transmitted by respiratory secretions (cough, snee

Describe insulin resistance in non-conventional factors, Describe Insulin r...

Describe Insulin resistance in Non-conventional Factors ? Insulin resistance refers to a generalized metabolic disorder in which various tissues are resistant to normal levels of

Explain phylum coelentrat, Phylum Coelentrat a : (9,000 'species) The ...

Phylum Coelentrat a : (9,000 'species) The body is radial and made up of two cell layers. Gastro-Vascular cavity is saclike, tentacles and stinging cells are present. They may

Diabetes mellitus, 1. How often should she have Glycated hemoglobin (HgbA1C...

1. How often should she have Glycated hemoglobin (HgbA1C) drawn?  What percentage is desired? 2. How often should she see an ophthalmologist or optometrist? 3. What should be

Define important points while working with autoclave, Define Important Poin...

Define Important Points While Working With Autoclave? Note: Following points should be kept in mind while working with autoclave. 1. Autoclave should not be packed tightly o

Explain the concepts of chromosome, How are the concepts of chromosome, chr...

How are the concepts of chromosome, chromatin and chromatids related? In which phase of the cell cycle does DNA duplicate? Chromatin is a set of filamentous DNA molecules dispe

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