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

Digestive system - large intestine, LARG E INTESTINE - More wider. Arr...

LARG E INTESTINE - More wider. Arranged around small intestine in the form of ? . It is 1.5 m long. Villi & microvilli absent. It shows 3 parts Ceacum, colon & rectum.

Define treatment and management of adverse food reactions, Define Treatment...

Define Treatment and Management of Adverse Food Reactions? Management of adverse food reactions involves diagnosing the problem followed by nutritional and medical care.

Arthropod classes characterized to the body division, Q. How are the three ...

Q. How are the three major arthropod classes characterized according to the body division? In arachnids and crustaceans the head is fused with the thorax forming the cephalotho

Explain trapezoidal full mucoperiosteal flaps, Explain Trapezoidal Full Muc...

Explain Trapezoidal Full Mucoperiosteal Flaps a) Similar to rectangular flap except: The 2 vertical incisions are at obtuse angle with the Horizontal incision, b) Mostly

Tolerance range - ecosystem, Tolerance Range - Ecosystem Organisms are...

Tolerance Range - Ecosystem Organisms are able to survive only within certain maximum and minimum limits with respect to each environmental factor such as water, light and tem

Asexual reproduction and cloning in plants, before stem cuttings are plante...

before stem cuttings are planted the cut end of the stem is often dipped in a hormone powder .what is the point of this?

Onion cells, What is the process of mitosis in onion root tips in a lab

What is the process of mitosis in onion root tips in a lab

Describe the initiation of visual impulse, Describe the Initiation of Visua...

Describe the Initiation of Visual Impulse When light falls on the retina it is absorbed by the photosensitive pigments of rods (rhodopsin) and cones (photopsin). It initiates

LPN, In many tests it is acceptable to read a positive result before the in...

In many tests it is acceptable to read a positive result before the incubation time is completed. why is this not the case with starch agar?

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