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

What are the two divisions of the autonomic nervous system, Q. What are the...

Q. What are the two divisions of the autonomic nervous system? The autonomic nervous system is divided into the parasympathetic nervous system and the sympathetic nervous syste

Does ph affect the enzyme activity, Q. Does pH affect the enzyme activity? ...

Q. Does pH affect the enzyme activity? The concentrations of hydrogen ions in solution affect the enzyme activity. Each enzyme has utmost efficiency under an optimum p H . S

Becoming online tutor, how can i work with u as an online biology tutor?

how can i work with u as an online biology tutor?

Statistical measures - population, Statistical Measures - Population A...

Statistical Measures - Population A population has a number of group characteristics that are statistical measures unique to the population group and are not the characteristi

History of evolution, History of evolution? Evolution is usually define...

History of evolution? Evolution is usually defined as "change over time." In spite of the incredible diversity of life found on Earth, many fundamental characteristics are shar

Splenic abscess, Splenic infarction is a common complication of left-sided ...

Splenic infarction is a common complication of left-sided IE (40 per cent of cases). Only 5 per cent of patients with splenic infarction will develop splenic abscess. This infectio

What happens to the movement of molecules at equilibrium, What happens to t...

What happens to the movement of molecules at equilibrium? At equilibrium, the movement of molecules continues, but because there is no concentration gradient, there is no net m

What conclusion can draw about microorganisms, What conclusion you can draw...

What conclusion you can draw about microorganisms that you saw under microscope from the plaque of your teeth?

Show coding for stimulus intensity, Q. Show Coding for stimulus intensity? ...

Q. Show Coding for stimulus intensity? Coding for stimulus intensity: If AP's are all-or-none responses that do not vary in amplitude, how does the NS distinguish between stim

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