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

Explain the lectins or haemagglutinins, Explain the Lectins or Haemagglutin...

Explain the Lectins or Haemagglutinins? These are sugar binding proteins, having the ability to bind and agglutinate red blood cells (RBCs). You will find that they occur in mo

Which of groups is not ionizable, Which of the following groups is NOT ioni...

Which of the following groups is NOT ionizable? Select one: a. Guanidinium b. Imidazole c. Phosphoryl d. Amine e. Aldehyde

Respiratory lining of human lung, Normal 0 false false fals...

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

How we can do karyotype analysis, Why in the preparation of a karyotype ana...

Why in the preparation of a karyotype analysis is the use of a substance like colchicine interesting? Colchicine is a substance that disallows the formation of microtubules and

What are the cells that produce the myelin sheath, What are the cells that ...

What are the cells that produce the myelin sheath? Of which substance is the myelin sheath formed? In the central nervous system (CNS) the myelin sheath is made by apposition o

Triple vessel disease , Triple Vessel Disease (TVD) :  Patients with tripl...

Triple Vessel Disease (TVD) :  Patients with triple vessel disease and impaired left ventricular function do badly on medical treatment. They are candidates for CABG. Operation is

What is the greenhouse effect, What is the greenhouse effect? The green...

What is the greenhouse effect? The greenhouse effect is a phenomenon in which atmospheric gases like CO 2 trap reradiated heat from the Earth, much as the glass panes of a gre

What are the symptoms of congestive cardiac failure, Q. What are the sympto...

Q. What are the symptoms of congestive cardiac failure? Congestive cardiac failure is a progressive form of cardiomyopathy. The most classical symptom is fluid imbalance due to

Planting bacteria gardens, Planting bacteria gardens When the bacteria ...

Planting bacteria gardens When the bacteria gardens have cooled place them out on the table but do not increase the glass covers unless you are ready to plant them. Toothpicks

Define skirt fold thickness (spt) method, Define Skirt Fold Thickness (S...

Define Skirt Fold Thickness (SPT) Method? Skin fold measurement is the most widely used field method of body composition assessment. The skin fold (SKF) is an indirect measu

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