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

Explain the food applications of carrageenan, Food Applications of Carragee...

Food Applications of Carrageenan Carrageenan consists of a family of hydrocolloids, which have different properties and it has a wide variety of uses. Some examples of properti

#title, natural selection

natural selection

Cardio pulmonary resuscitation, CARDIO PULMONARY RESUSCITATION: Cessat...

CARDIO PULMONARY RESUSCITATION: Cessation of cardiac activity is determined by  inability to palpate a central pulse,  unresponsiveness and Gnea. CPR consists of  measures for

Importance of care in children, Importance of care in children Childre...

Importance of care in children Children  are not  just  small adults. This is important  to keep  in mind when evaluating how children react to illness, perceive an illness, o

What is oral hygiene, What is Oral Hygiene  An index can be used to eva...

What is Oral Hygiene  An index can be used to evaluate the oral hygiene level in the patient e.g. Oral hygiene Index (Green and vermilion) or Plaque index (Silness and Loe). An

What is the meaning of accumulation, What is the meaning of accumulation ...

What is the meaning of accumulation The word "accumulation" is used in many of the definitions in the sense that th e horizon must have more of the material in question than i

Ground water sources and characteristics, After glaciers, ice caps and snow...

After glaciers, ice caps and snowfields, ground water is the next largest fresh water reservoir. Precipitation that does not evaporate back into the air and does not run off over t

What is initiation delay, Which of the following is the best definition of ...

Which of the following is the best definition of the initiation delay that is seen in all organisms? A. Initiation delay is explained as the time it takes for the pre-initiatio

Potassium (k) - macronutrients, Potassium (K) - Macronutrients The mai...

Potassium (K) - Macronutrients The main source of K + for plants comes from withering of K containing minerals. Potassium released by withering dissolves in the soil solution

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