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

Why dairy or milk product are important for human body, Why Dairy or milk p...

Why Dairy or milk product are important for human body? This first food of mammals is rich in body-building proteins and bone-forming calcium, besides being the only source of

Development of fruit, Development of Fruit Concurrent with the develo...

Development of Fruit Concurrent with the development of the seed(s), the ovary is transformed into a fruit. The fruit protects the seeds and allows their release or germinati

Explain gastric bypass - surgical management for obesity, Explain Gastric B...

Explain Gastric Bypass - surgical management for obesity? Surgery is the current 'gold standard' for bariatric surgical procedures. It involves use of a stapling device to crea

Deficiency diseases-copper deficiency, Copper deficiency Copper deficie...

Copper deficiency Copper deficiency may be primary or secondary (conditional). Primary copper deficiency occurs due to its inadequate dietary intake; whereas secondary copper d

Illustrate the improper implant design, Improper Implant Design Out of ...

Improper Implant Design Out of the plethora of implant systems available it is the responsibility of the clinician to select the most suitable. In comparison of the straight cy

Explain the classification of hydrocolloids by function, Classification of ...

Classification of hydrocolloids by function It must be clear to you now that gum ghatti, gum karaya, gum tragacanth ( classified as exudate gums) locust bean  gum, guar g

Explain brotherhood and sisterhood using abo blood typing, Is it possible t...

Is it possible to perform investigation of natural paternity, maternity or brotherhood and sisterhood using the ABO blood typing? By using an ABO blood typing it is possible on

Disorders of motor neurons and the spinal cord, Disorders of motor neurons ...

Disorders of motor neurons and the spinal cord A number of movement disorders are produced by damage either to the spinal cord or to cortical projections to the spinal cord. Th

Explain the ps in vcd in cyanotic heart disease, Explain the PS In VCD in C...

Explain the PS In VCD in Cyanotic Heart Disease ? These are peculiar to this group only. It occurs in young infants (usually after two months and rarely after two years), It co

Nervous system - spinal nerves, SPINAL NERVES 31 Pairs. Mixed type. ...

SPINAL NERVES 31 Pairs. Mixed type. Total weight 150 gms. At the base of origin of spinal nerves gland of swammardon or calcarious ganglion is present. Composed of m

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