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

Solar tracking - nastic and epinastic responses, Solar Tracking - Nastic an...

Solar Tracking - Nastic and Epinastic Responses Many plants such as sun flowers are capable of solar tracking in which the flat blade of leaves or inflorescence will remain a

Agro industrial-proteins, Proteins Protease inhibitors: Protease inh...

Proteins Protease inhibitors: Protease inhibitors are protein-based substances widely distributed within the plant kingdom, including the seeds of most cultivated legumes wh

Explain assessment of iron status - serum ferritin, Explain assessment of i...

Explain assessment of iron status - Serum Ferritin? Serum Ferritin: This method is indicative of iron stores. As we know, a long term negative iron balance first results in dep

Preformation theory - pre-mendelian theory, Preformatio n Theory (1632-172...

Preformatio n Theory (1632-1723) (i)         Scientist 'Malpighi Swammerdam' propounded the preformation theory. This theory holds that the sex cells (sperm and ova) had the m

In which phase mother exchanges substances to fetus, Exchange of oxygen, nu...

Exchange of oxygen, nutrients, carbon dioxide and waste products from mother to fetus takes place through diffusion across the: a) Chorionic villi (pron: core-e-on-ick villi)

Find human anatomy & physiology laboratory manual, I can not find Human Ana...

I can not find Human Anatomy & Physiology Laboratory manual 10th edition cat version by Marieb/mitchell in expertsmind homework help, I have the text book just need to make sure I

Explain minerals requirement during thyphoid, Explain Minerals requirement ...

Explain Minerals requirement during thyphoid Minerals:   Loss of electrolytes and water is observed due to diarrhoea. Thus liberalizing on sodium intake  through salty soups,

Platyhelminthes, tell me the general characters of monopisthocotylea

tell me the general characters of monopisthocotylea

Importance of human milk for infant growth and development, Define the Impo...

Define the Importance of Human Milk for Infant Growth and Development? Let us now look at the value of human milk in promoting infant growth and development.  Success of lactat

What are the plant hormones, What are the plant hormones? The Plant hor...

What are the plant hormones? The Plant hormones also called as phytohormones, are the substances that control the embryonic development and the growth of the adult plant.

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