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

How nutritional status affects the bmr, How Nutritional status affects the ...

How Nutritional status affects the BMR? Under nutrition and starvation are the factors which require consideration. Prolonged under nutrition or starvation causes a reduction o

Explain tb in pregnancy, TB in pregnancy Treatment of TB should be init...

TB in pregnancy Treatment of TB should be initiated in pregnancy when there is moderate to high suspicion of disease because active infection during pregnancy poses a risk to t

What are the three main parts of a eukaryotic cell, The eukaryotic cell can...

The eukaryotic cell can be separated into two major portions: the cell membrane that divides the intracellular space from the outer space phisically delimiting the cell; the cytopl

What do you mean by zoological nomenclature, What do you mean by Zoological...

What do you mean by Zoological Nomenclature? This trust is housed in the British Museum, London and is responsible for some of the most important literature in Zoological Nomen

Drugs for sexually transmitted infections, Drugs for Sexually Transmitted I...

Drugs for Sexually Transmitted Infections Many infections can be transmitted during sexual contact. The text and tables that follow are limited to management of sexually transm

Trypsin, Trypsin Trypsin is secreted by the pancreas in an inactive fo...

Trypsin Trypsin is secreted by the pancreas in an inactive form trypsinogen. It is activated by enterokinase secreted by the glands in the intestinal wall. As trypsin is forme

Explain some do's and don'ts when working in laboratory, Explain some Do's ...

Explain some Do's and Don'ts when working in Laboratory? 1) Always wear a lab coat when working a laboratory. 2) Ensure that no harm is caused to yourself or the people work

The osmolarity of the rbcs and plasma after equilibration, Ed Rivers, a 3rd...

Ed Rivers, a 3rd year medical student, was alone in the hospital ER one night. It was unusually quiet that night, and the resident was getting some much needed sleep. A patient, Mr

Production of disease-free plants, Production of Disease-free Plants U...

Production of Disease-free Plants Under normal conditions plants are infected by a wide range of pathogens such as bacteria, fungi, viruses, viroids, and insects like nematode

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