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

How many are the pulmonary veins, Q. What and how many are the pulmonary ve...

Q. What and how many are the pulmonary veins? The pulmonary veins are part of the pulmonary circulation they are vessels that carry oxygen-rich (arterial) blood from the lungs

Show the characteristics of honey, Honey comes under the purview of Prevent...

Honey comes under the purview of Prevention of Food Adulteration Act (PFA). Due to its limited production and high cost, honey is prone to adulteration by cane sugar, invert sy

Define human digestive system, Q. Which organs of the body is part of the h...

Q. Which organs of the body is part of the human digestive system? The digestive system also known as "systema digestorium" or gastrointestinal system is composed of the digest

Camptotheca acuminata, This species is an ornamental tree in China, and has...

This species is an ornamental tree in China, and has yielded the clinically active agents topotecan, irinotecan, and 9 -aminocamptothecin, which are semisynthetically derived from

45, What grade did you get in biology?

What grade did you get in biology?

Explain the natural history of coronary artery diseases, Explain the NATURA...

Explain the NATURAL HISTORY OF CORONARY ARTERY DISEASES (GAD)? The natural history of CAD is very important from the preventive point or view. Though the usual manifestations o

Animal nutrition, briefly explain the following feeding mechanisms in holoz...

briefly explain the following feeding mechanisms in holozoic animals a) filter feeding b) fluid feeding c)deposit feeding

Gonial apospory and somatic apospory, Gonial Apospory and Somatic Apospory ...

Gonial Apospory and Somatic Apospory Gonial Apospory There is no meiosis. The megaspore mother cell enlarges; a small vacuole appears above and below the nucleus. The c

Determine the food sources of folate, Determine the Food Sources of Folate?...

Determine the Food Sources of Folate? Folate occurs naturally in foods. Although folate is found in a wide variety of foods, it is present in a relatively low density except i

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