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

Microbiology, 2. Vaccinations against various childhood diseases have contr...

2. Vaccinations against various childhood diseases have contributed to the entry of women, particularly mothers, into the fulltime workplace. a. Is this statement supported by data

Explain theory or principle of determination of coliforms, Explain Theory o...

Explain Theory or Principle of Determination of Coliforms? Coliforms are gram negative, non-spore forming, facultative anaerobic bacteria which produce acid and gas on lactose

Mitral valve replacement with allograft , Mitral Valve replacement with All...

Mitral Valve replacement with Allograft (Mitral homograft) :  C. Acar and colleagues used mitral allograft for mitral valvc replacement using a new technique of fixation of p

How auxins act helping the lateral growth of the stem, How auxins act helpi...

How auxins act helping the lateral (secondary) growth of the stem? The Auxins stimulate the formation of conductive vessels, phloem and xylem, promoting the thickening of the s

Define advantages of using bacteria as a source of protein, Define Advantag...

Define Advantages of using Bacteria as a source of protein? Advantages of using Bacteria as a source of protein are as follow: 1. High yield of protein (60-80%) 2.

Define requirement of energy for physical activity, Define requirement of e...

Define requirement of energy for Physical activity? This is the most variable and, after BMR, the second largest component of daily energy expenditure. Humans perform obligator

What are the basic constituents of the cell membrane, What are the basic co...

What are the basic constituents of the cell membrane? The cell membrane is produced of lipids, proteins and carbohydrates. The membrane lipids are phospholipids, a special t

Determine the term - angiomas, Determine the term - Angiomas Angiomas a...

Determine the term - Angiomas Angiomas are congenital collections of abnormal vessels that divert the normal flow of blood. These capillaries, venous, or arteriovenous (A-V) ma

Management of soil productivity, Management of soil productivity The co...

Management of soil productivity The continuous prosperity and well being of the people of any nation is dependent upon several factors, one of the most important being the leve

Ribosomes, Ribosomes - It is also known as Protein factory, Engine ...

Ribosomes - It is also known as Protein factory, Engine of the cell. Ribosomes are smallest cell organelles (150 x 250 Å). Unit membrane less cell organelle. Ribo

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