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

Define the operations of a public nutritionist, Define the Operations of a ...

Define the Operations of a Public Nutritionist? A public nutritionist can perform the following: In the hospital-based set up, she is a part of the team delivering thera

Anticodon, Anticodon is the series of three nucleotides on the transfer RN...

Anticodon is the series of three nucleotides on the transfer RNA molecule which recognizes and pairs with the specific codon on a messenger RNA molecule; it helps in controlling t

Explain risk assessment, Explain Risk Assessment Risk Assessment   :  ...

Explain Risk Assessment Risk Assessment   :  The scientific evaluation  of  known  or  potential adverse  health effects  resulting  from  human exposure to food borne hazards

Phylum platyhelminhes, what are the classification of phylum platyhelminthe...

what are the classification of phylum platyhelminthes?

Ichnology, Ichnology :  This is the study of foot prints. Ichnology is a ty...

Ichnology :  This is the study of foot prints. Ichnology is a type of branch of geology which deals with traces of organismal behavior like as burrows or footprints. It is commonly

Plants and animals - biological weathering, Plants and animals The acti...

Plants and animals The activity of living organisms like lichens, fungi, bacteria, blue green algae, bryophytes is also responsible for weathering.  It is referred to as biolog

Explain the proteus - characteristics of bacteria, Explain the Proteus - Ch...

Explain the Proteus - Characteristics of Bacteria? It is gram negative, non-sporulating rod, which is characterized by rapid motility (peritrichous flagella) and swarming type

Explain food lipids, Explain Food lipids It is either consumed in the ...

Explain Food lipids It is either consumed in the form of "visible" fats, which have been separated from the original plant or animal sources, such as vegetable oil and butter,

Pesticides - danger to wildlife, Pesticides - Danger to Wildlife Anoth...

Pesticides - Danger to Wildlife Another recently realised danger to wildlife in many parts of the world has come from the development of more effective pesticides. As agricult

Describe the function of gastrin in the digestive process, Q.How is it prod...

Q.How is it produced and what is the function of gastrin in the digestive process? The existence of food in the stomach stimulates the secretion of gastrin that in its turn tri

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