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

Draw a diagram of transverse section of mammalian bone, Draw a diagram of t...

Draw a diagram of transverse section of mammalian bone and label the following parts in it: (i) Haversian canal (ii) Lacuna containing osteocytes (iii) Canaliculi

Explain the general characteristics of honey, A) General Characteristics ...

A) General Characteristics 1) It should be a well-ripened natural product. 2) It should be clear and visually transparent. 3) It should be extracted with the help of a

Which term refers to muscles with less than normal tone, In biological stud...

In biological studies which of the below terms is used to refer to muscles with less than normal tone? Is it: a) Latent b) Atonic c) Flaccid d) None of these ANSWE

Mitosis, what is mitotic apparatus

what is mitotic apparatus

Water cycle-hydrological cycle, Water Cycle (Hydrological Cycle) The mo...

Water Cycle (Hydrological Cycle) The movement of water on the earth is continuous and forms many complex inter-related loops (Figure shown below). Cycling of water involves atm

What is the principle of the design, In designing an experiment to find out...

In designing an experiment to find out whether light is required for photosynthesis    (a) what is the principle of the design    (b) what control would you use?

Explain capillaries of the vascular system, Q. What are the capillaries of ...

Q. What are the capillaries of the vascular system? Capillaries are small blood vessels that perform exchange of substances between the tissues and blood the body Capillaries a

Observations reveal about mechanisms of sex determination, XYY humans are f...

XYY humans are fertile males. XXX humans are fertile females. What do these observations reveal about the mechanisms of sex determination and dosage compensation?

Define metabolic aberrations and clinical manifestations, Define Metabolic ...

Define Metabolic Aberrations and Clinical Manifestations? The state of obesity brings about certain alterations in the normal body processes which are enumerated herewith and h

Why soybean protein used as stabilized dispersions, Soybean protein used as...

Soybean protein used as stabilized dispersions Soybean protein concentrates have been used as stabilized dispersions in milk-like beverages and simulated dairy products, such a

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