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

State about osteogenesis, State about osteogenesis Embryologically, ost...

State about osteogenesis Embryologically, osteogenesis may be classified as either intramembranous or endochondral. When the ossification occurs directly, it is defined as intr

Wbc.., Ask question #Minimuwwm 100 words accepted#

Ask question #Minimuwwm 100 words accepted#

Role of private sector in health care, Role of Private Sector in Health Car...

Role of Private Sector in Health Care One of the biggest policy challenges for the government is to decide on the appropriate areas and a suitable policy framework for having

Financial accounting software, Financial Accounting Software An Ide...

Financial Accounting Software An Ideal Accounting Software should have the following features:  Purchase Details  Invoice Details  Purchase/Sale Return (Debit/Cre

Strategies for help of diabetic patients, It is important that when patient...

It is important that when patients change behaviour they expectt to see results quickly; therefore, the new behaviour must have advantages. So someone who wants to control blood su

Respiration, what is the organ of respiration in snaks

what is the organ of respiration in snaks

Botany, how can i earn money by submitting assignments?

how can i earn money by submitting assignments?

Shifting attention by dopamine, Q. Shifting attention by Dopamine? Dopa...

Q. Shifting attention by Dopamine? Dopamine: Dopamine plays a pivotal role in aspects of shifting attention. Administration of D1/D2 receptor antagonist haloperidol impairs the

Explain about dry heat sterilization, Q. Explain about Dry heat sterilizati...

Q. Explain about Dry heat sterilization? It is used to sterilize glass ware, forceps, scissors, scalpels, all glass syringes, swabs, some pharmaceuticals products such as liqui

Disorders related to nutrition and digestive system, DISORDE R RELATED TO ...

DISORDE R RELATED TO NUTRITION AND DIGESTIVE SYSTEM Malnutrition Malnutrition is improper intake of food. Malnutrion covers both undernutrition and overnutrition. Und

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