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

Define the activation of an agonist during movement, Explain the relationsh...

Explain the relationship between an agonist muscle and its antagonist as it relates to positional control, functional movement and neuromuscular activation. For example, how the th

What is genetic mapping, How can the concept of recombination frequency be ...

How can the concept of recombination frequency be used in genetic mapping? Genetic mapping is the determination of the location of the genes in a chromosome. By determining

Membrane proteins and carbohydrate, The cell membrane is a slim semi-permea...

The cell membrane is a slim semi-permeable membrane which surroundings the cytoplasm of a cell. Membrane function is to defend the integrity of the interior of the cell by allowing

Explain some ethological activities, Explain Some Ethological Activities ...

Explain Some Ethological Activities Comparative studies of behaviour or activities are also used in systematic for identification of organisms. The comparative study of the beh

Determine some food sources for copper, Determine some Food Sources for Cop...

Determine some Food Sources for Copper? Foods containing more than 1 mg copper per 1000 kilocalories are considered high in copper and include green leafy vegetables, nuts, leg

Explai total fertility rate and replacement level, Differentiate between To...

Differentiate between Total Fertility Rate (TFR) and Replacement Level (RL) Due to uncontrolled excessive hunting the population of tigers in a forest becomes Zero. Discuss the

Viruses, Viruses Viruses are sub cellular, ultramicroscopic infectious...

Viruses Viruses are sub cellular, ultramicroscopic infectious agents of 20 nm-300 nm size range Many of them cause diseases like polio, dengue, small pox, measles, rabies, com

Define oxidised starch, Oxidised starch Oxidised starch finds a number ...

Oxidised starch Oxidised starch finds a number of uses in the food industry where a neutral tasting, low viscosity 'body builder' is required as in lemon curd manufacture, in s

Survival of the saddleback, The North Island saddleback or tieke, Philestur...

The North Island saddleback or tieke, Philesturnus rufusater, is an endemic New Zealand bird. Saddlebacks inhabit the middle or lower layers of the forest, and eat small invertebra

Codeine, CODEIN E - It is methylmorphine, obtained from opium. I...

CODEIN E - It is methylmorphine, obtained from opium. It is used in cough syrups. A notable side effect of codeine is constipation.

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