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

Explaintransketolase, ExplainTransketolase Transketolase with the help...

ExplainTransketolase Transketolase with the help of TPP  and Mg ++   is  required again. This  time  it  transfers carbon 1+2 from xylulose-5-phosphate  to erythrose-4-phospha

What are the vascular bundles, What are the vascular bundles? How does conf...

What are the vascular bundles? How does configuration of the vascular bundles within the stem differentiate monocots from dicots? The Vascular bundles are segments of xylem and

Osmotic strength of the fluids, Osmotic Strength of the Fluids in Immediate...

Osmotic Strength of the Fluids in Immediate Surrounding The availability of soluble mineral salts varies widely from habitat to habitat. In fact, a big chunk of land in our

Define the skin capillary fragility test, Define the Skin Capillary Fragili...

Define the Skin Capillary Fragility Test? One of the earliest signs of ascorbic acid deficiency is the fragility of the capillaries. Capillary fragility is measured by applyin

Perceptual disorders, Perceptual Disorders The procedure actually const...

Perceptual Disorders The procedure actually constitute a sub-battery and include tests of the subject's ability to recognise shapes by touch and identifies numbers written on t

What is green fluorescent protein, Q. What is green fluorescent protein (GF...

Q. What is green fluorescent protein (GFP)? Green fluorescent protein is a protein which is found in a luminescent jellyfish (Aquorea victoria) that lives in cold waters of nor

Carbohydrates requirements for ulcerative colitis, Q. Carbohydrates require...

Q. Carbohydrates requirements for ulcerative colitis? Carbohydrates: They form the easily absorbable source of energy. Bulk-producing vegetables are restricted so as to allow b

What are nadph and nadp, Q. What are NADPH and NADP? NADP is the short ...

Q. What are NADPH and NADP? NADP is the short form of the nicotinamide adenine dinucleotide phosphate cation, a hydrogen acceptor. NADPH is made when NADP binds to one hydrogen

What is oxidative phosphorylation, What is oxidative phosphorylation? O...

What is oxidative phosphorylation? Oxidative phosphorylation is  the process by which ADP is phosphorylated by Pi  to ATP  in  the  respiratory chain.

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