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

Neuropsychological understanding of behavioural deficits, Neuropsychologica...

Neuropsychological understanding of behavioural deficits Behaviour is an outcome of the interaction of the brain with the environment. A composite of multiple psychological pro

Bacterial diseases-sheep and goats, Sheep and goats The disease is rea...

Sheep and goats The disease is readily transmissible to human being from goats, and is known as Malta fever, Mediterranean fever or undulant fever. The chief causal agent is B

What is atrial switches operations, What is Atrial Switches Operations (Sen...

What is Atrial Switches Operations (Senning or Mustard Operation) ? In transposition of the great arteries there is ventriculo arterial discordance whereby aorta arises from ri

Specific causes of parental anxiety - hospitalization, Specific Causes of P...

Specific Causes of Parental Anxiety   Some of the factors  that increase  their anxiety include following:  1)  Fear of the strange environment in the hospital  2)  Fear

Fowl cholera, F o wl cholera Fowl cholera, a highly contagious diseas...

F o wl cholera Fowl cholera, a highly contagious disease of poultry caused by Pasteurella multocida, was one of the first infectious diseases to be recognized by Louis Past

Aerobic respiration, Ask question end product of aerobic repiration ar...

Ask question end product of aerobic repiration are

Health impacts of water pollution, Health Impacts of Water Pollution I...

Health Impacts of Water Pollution It is well known that clean water is absolutely essential for healthy living. Although adequate supply of fresh and clean drinking water is a

Sour taste, Sour taste is usually due to the presence of acids such as acet...

Sour taste is usually due to the presence of acids such as acetic acid, citric acid, benzoic acid. Upon dissociation, the acids give H ions which impart acidity. More the H ion con

Name an artificial fertiliser which farmers can use, Name an artificial fer...

Name an artificial fertiliser or fertilisers which farmers can use to increase the supply of nitrate, phosphate and potassium to their crops.   NPK compound fertiliser havin

Explain taxonomy - modern trends in animal taxonomy, Explain Taxonomy - Mod...

Explain Taxonomy - Modern Trends in Animal Taxonomy The term taxonomy which is derived from the Greek word 'taxis', meaning order or arrangement and nbmos ' meaning law, deals

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