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

Coding sequence, Coding sequence is the portion of the gene or the mRNA wh...

Coding sequence is the portion of the gene or the mRNA which actually codes for a protein. Introns are not coding for the sequences; nor are the 5' or 3' untranslated regions (or

What are the cnidocytes, Q. What are the cnidocytes? What is the name of th...

Q. What are the cnidocytes? What is the name of the capsule inside the cnidocyte? What are the biological functions of this structure? Cnidocytes are specific cells present in

Explain measles, Measles Adults born after 1956 who have not received ...

Measles Adults born after 1956 who have not received 2 doses of live measles vaccine (not the killed vaccine that was commonly used in the 1960s) after their first birthday an

Procedures related to hygenic care, PROCEDURES RELATED TO HYGENIC CARE ...

PROCEDURES RELATED TO HYGENIC CARE Would you like to remain without a bath on any given day? Do you remain without a bath when you are sick.? No one normally, would like to m

Duck virus hepatitis, Duck virus hepatitis This highly contagious disea...

Duck virus hepatitis This highly contagious disease of ducks is caused by a member virus belonging to genus Enterovirus of the family Picornaviridae. It mostly affects duckling

What are the factors that for influencing photosynthesis, Q. What are the f...

Q. What are the factors that for influencing photosynthesis also interfere with the gross primary productivity? Mostly light and water, but as well mineral salts, carbon dioxid

Disaster, The Word 'disaster' is believed to be originating from the French...

The Word 'disaster' is believed to be originating from the French word 'Disastre' which is a combination of two words 'des' meaning bad and 'astre' meaning star. Thus this term ref

Calculate temperature for survival & growth of microorganism, Calculate Tem...

Calculate Temperature for Survival and Growth of Microorganisms? Survival and growth of microorganisms is also affected by the temperature. According to their temperature requi

What is bioremediation, What is bioremediation? Bioremediation is the u...

What is bioremediation? Bioremediation is the use of microorganisms, as bacteria, protists and fungi, to degrade noxious substances turning them into non toxic or less toxic su

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