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 is the meaning of cardiovascular system , What is the meaning of Cardi...

What is the meaning of Cardiovascular System ? The cardiovascular system takes care of the distribution of gases, nutrients, hormones, immune elements, and the removal of waste

Are the extraembryonic membranes the same in all vertebrates, Q. Are the ex...

Q. Are the extraembryonic membranes the same in all vertebrates? The presence of each extraembryonic membrane differs according to the vertebrate class. In amphibians and fi

Homogeneity to the nucleic acid chains, Q. Do the phosphate and the pentose...

Q. Do the phosphate and the pentose groups give heterogeneity or homogeneity to the nucleic acid chains? What about the nitrogen- containing groups? Supported by that, which of tho

Phylogeny and evolution, While studying evolution, a student comes across a...

While studying evolution, a student comes across a cladogram that includes clades like amphibia, reptilia, aves, and mammalia. What must be the basal clade?

Procedure of the second interphase period, Q. What are the events that mark...

Q. What are the events that mark the end and the beginning of the second interphase period and what happens in the cell in this period? The second interphase period is the S, i

What is ecology, What is Ecology? Ecology is the field of Biology that ...

What is Ecology? Ecology is the field of Biology that studies the relationship among living beings and among living beings and the environment.

Pumps, The pumps The blood from the operating field is sucked with t...

The pumps The blood from the operating field is sucked with the help of the cardiotomy sucker and pump, which is filtered to remove all particulate matter and then pass

Eye drops, Eye Drops Medication is instilled in the form of drops or ...

Eye Drops Medication is instilled in the form of drops or ointment.   Purpose To dilate pupil e.g. Atropine 1 per cent, drosyn 5 per cent or 10 per cent  To c

What mechanism of regulation utilized in maintaining, What mechanism of reg...

What mechanism of regulation is utilized in maintaining homeostatic balance of sugar?

Goal of bcc programme, The overall goal of BCC programs for diabetes mellit...

The overall goal of BCC programs for diabetes mellitus is to promote behaviors that control diabetes mellitus and prevent complications. These include: Following treatment

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