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

Skeletal system - hyoid bone, HYOI D BONE - Also called tongue bone...

HYOI D BONE - Also called tongue bone as lies at the base of the tonuge. One in number. Muscles of tongue and throat are attached to it. Not e :- Except

How is price mechanism or supply and demand concerned, How is price mechani...

How is price mechanism or supply and demand concerned?   The price mechanism or supply and demand: The price mechanism or supply and demand is concerned with how buyers

Favus, Favus Fowl favus is a superficial, chronic, sporadic mycotic in...

Favus Fowl favus is a superficial, chronic, sporadic mycotic infection of birds caused mainly by Trichophyton gallinae. Very rarely Trichophyton simii is also involved in the

What is the concept of universality of the genetic code, What is the concep...

What is the concept of universality of the genetic code? What are the exceptions to this universality? The genetic code is universal because the rules of protein codification b

What is haccp, What  is  HACCP? HACCP, as you may  already know,  is ...

What  is  HACCP? HACCP, as you may  already know,  is an acronym that stands for Hazard Analysis Critical  Control Point,  a systematic, science-based approach  used  in  food

Diabetes mellitus, 1. How often should she have Glycated hemoglobin (HgbA1C...

1. How often should she have Glycated hemoglobin (HgbA1C) drawn?  What percentage is desired? 2. How often should she see an ophthalmologist or optometrist? 3. What should be

Explain tertiary and quaternary structure of proteins, Explain Tertiary str...

Explain Tertiary structure and Quaternary structure of proteins Tertiary structure Secondary structure, in turn, folds back and bonds to itself in a three-dimensional mann

Human reproduction, Describe the events which lead to the formation of (a) ...

Describe the events which lead to the formation of (a) identical twins, (b) fraternal twins in detail

Precautions for selective and differential medium, Precautions for Preparat...

Precautions for Preparation of Selective and Differential Medium 1. Dissolve ingredients one by one. 2. Check and set the medium to appropriate pH. 3. Autoclave the mediu

Natural disasters, EARTHQUAKES          An earthquake with its terrible...

EARTHQUAKES          An earthquake with its terrible aftereffects is one of the most frightening and destructive phenomena of nature. It can be defined as a sudden movement of

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