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

Avian leukosis (sarcoma group of retroviruses), A v i a n leukosis (Sar...

A v i a n leukosis (Sarcoma group of retroviruses) This is a complex of viral diseases caused by an avian retrovirus with various manifestations such as lymphoid leukosis,

Define the energy requirement to avoid underweight problem, Define the Ener...

Define the Energy requirement to avoid Underweight problem? The total calorie intake should be 500 to 1000 Kcal in excess of the daily needs in order to result a gain in weight

Define in brief about the term - mri, Define in brief about the term - MRI ...

Define in brief about the term - MRI MRI is a more recent development that was initially introduced as a rival to CT. The technique itself is complex, relying on measurement o

What are homologous chromosomes, What are homologous chromosomes? Which are...

What are homologous chromosomes? Which are the human cells that do not have homologous chromosomes? Chromosomes have genes (genetic information in the form of nucleotide sequen

Demonstrate the structure of mitochondria, Demonstrate the structure of mit...

Demonstrate the structure of mitochondria The structure of mitochondria has been examined in great detail with the aid of the electron microscope, and it is now possible to des

Respiratory system, describe the structure of a lung in terms of covering m...

describe the structure of a lung in terms of covering membranes and cavities

Explain oncogenes or defective tumour suppressor genes, 1.  Explain why a g...

1.  Explain why a gain of function would be a dominant effect and a loss of function would be a recessive effect. Which types of genes are each associated with: oncogenes or defect

Explain adverse effects of fosamprenavir calcium, Explain adverse effects o...

Explain adverse effects of fosamprenavir calcium   Adverse effects are similar to those with amprenavir, but in clinical studies the incidence of nausea, vomiting and severe ra

Difference between rods and cones, Difference between rods and cones - ...

Difference between rods and cones - Rods Cones   1. More in number 1150 lakh   2. Outer seg. is cylindrical and contains

Rheumatic fever and rheumatic heart diseases, Rheumatic Fever and Rheumatic...

Rheumatic Fever and Rheumatic Heart Diseases Rheumatic fever is an inflammatory disease of the heart potentially involving all layers of the heart. The resulting damage to t

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