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

Proteins, what re conjugated proteins and how are they classified?

what re conjugated proteins and how are they classified?

Food processing and preservation, various changes that occur during concent...

various changes that occur during concentration of food

Goal of fostering the integration of biology, Evaluate how best to achieve ...

Evaluate how best to achieve the goal of fostering the integration of biology, statistics, and mathematics. What impediments stand in the way of progress? What is the best way to

Polypeptide chains of hemoglobin, Q. In sickle cell anemia, a hereditary di...

Q. In sickle cell anemia, a hereditary disease, there is replacement of one amino acid by another in one of the four polypeptide chains of hemoglobin. In this case are all of the s

Quantification of pulmonary stenosis, Quantification of Pulmonary Stenosis ...

Quantification of Pulmonary Stenosis :   Quantification by ECHO By Valve area Mild                 > 1.5 cm 2 Moderate         0.5-1.5 cm 2 Severe

Summer stratification - thermal stratification, Summer Stratification - The...

Summer Stratification - Thermal stratification Thermal stratification is fairly pronounced during the summer seasons in most lakes of the temperate (cold) regions but is rare

Where can rna are found within cells, Q. Where can RNA are found within cel...

Q. Where can RNA are found within cells? In the eukaryote cell nucleus the RNA can be found to dispersed in the nuclear fluid, along with the DNA, and as the main constituent o

What is the meaning of food exchanges, What is the meaning of Food Exchange...

What is the meaning of Food Exchanges Food exchanges mean grouping of foods in which the specified amount of all the food provides approximately equal amounts of carbohydrate,

What is the part of the female reproductive system, What is the part of the...

What is the part of the female reproductive system where fecundation occurs? Fecundation generally happens in the Fallopian tubes but it can also take place within the uterus.

Gametes, examples of gametes

examples of gametes

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