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

Vesicular stomatitis, Vesicular stomatitis Vesicular sto matitis virus ...

Vesicular stomatitis Vesicular sto matitis virus be longs to the ge nus Vesiculovirus in fa mily Rhabdoviridae. The disease clinically resembles foot-and-mouth disease, but is

Animals vs plants, Animals vs Plants Organisms are of two main types an...

Animals vs Plants Organisms are of two main types animals and plants, although all the above mentioned Unifying concepts of Biology apply equally to both animals and plants, ye

Osmoregulatory animal, Osmoregulatory Animal An osmoregulatory animal ...

Osmoregulatory Animal An osmoregulatory animal is generally in an osmotic steady state even though there may be hourly and daily variations in osmotic balance. The concentrati

Explain adverse effects of caspofungin, Explain Adverse Effects of Caspofun...

Explain Adverse Effects of Caspofungin Although generally well tolerated, caspofungin occasionally causes rash, fever and mild hepatic toxicity (Medical Letter 2001; 43:58). An

Determine about the nervous tissues, Determine about the nervous tissues ...

Determine about the nervous tissues The nervous tissues throughout the body, including the visual nervous system, are continuously active. Electrical reactions can only be indu

What is the importance of fungi, What is the Importance of Fungi? Fungi...

What is the Importance of Fungi? Fungi are widely distributed in nature. It is estimated that there are about 1.5 million species of fungi on earth. Fungi are both beneficial a

Intrisic and Extrinsic Limiting Factors, How do the ideas of energy and che...

How do the ideas of energy and chemical cycles, community structure, biodiversity and succession fit together to form the basis of the way the natural world works?

1, characters of phylum protozoa

characters of phylum protozoa

Define natural gums - hydrocolloids, Natural Gums These are derived fro...

Natural Gums These are derived from various plant as well as microbial sources and are divided into the following four basic categories: 1.  Seaweed gums:   Agar, Alginate,

Instrument examination and care, Instrument Examination and Care Cleani...

Instrument Examination and Care Cleaning instruments, provides a good opportunity to examine, replace or remove damaged instruments; lubricate items such as handpieces; and oth

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