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

Pollen tube structure, Pollen Tube Structure The pollen tube in the s...

Pollen Tube Structure The pollen tube in the stigma is filled with cytoplasm containing numerous mitochondria and dictyosomes. The number of dictyosome cisternae is reduced i

Strategic and tactic control of parasitic diseases, S t r a tegic and t...

S t r a tegic and tactic control of parasitic diseases The reduced performance of the animals in terms of decreased milk yield, draught power, breeding performance, poor bo

Abolition of arrhythmias by exercise, Q. Abolition of Arrhythmias by Exerci...

Q. Abolition of Arrhythmias by Exercise? The induction of arrhythmias by exercise is well recognized, but the abolition of ectopic activity is less commonly appreciated. The me

Demography, DEMOGRAPH Y - Scientific study of vital & social statistic...

DEMOGRAPH Y - Scientific study of vital & social statistics such as births, deaths, disease etc. of human population is called demography. It deals with - 1.      Change

Normal color vision- color blindness, If a color blind man marries a woman ...

If a color blind man marries a woman pure for normal color vision, it's probable that one of the below situations may result. Is it possible that a) All the children would be

Fensom and spanner electroosmotic flow hypothesis, Fensom and Spanner Elect...

Fensom and Spanner Electroosmotic Flow Hypothesis In electroosmosis the ions flow across a membrane in response to electrical gradient. Ions pull along water and other content

How does the female gamete move from the ovary to the uterus, Q. How does t...

Q. How does the female gamete move from the ovary to the uterus? The female gamete released from the ovary falls into the surrounding abdominal cavity and is collected by the F

Explain the kidney function in human biology, Explain the Kidney Function i...

Explain the Kidney Function in human biology? Blood first enters the capillaries in Bowman's capsule where it is filtered. The pores in the capillary walls allow water and smal

How delayed complementary feeding cause pem, How Delayed Complementary Feed...

How Delayed Complementary Feeding Cause PEM? The mothers from poorer socioeconomic groups where PEM is more prevalent, delayed introduction of complementary foods (foods in add

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