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 are the main structures of the human eye, What are the main structures...

What are the main structures of the human eye? The major structures of the human eye are the cornea, the iris, the pupil, the ciliary muscles, the crystalline lens and the ret

Parathyroid disorders, PARATHYROID DISORDERS - (i) Hypoparathyroidism ...

PARATHYROID DISORDERS - (i) Hypoparathyroidism (deficiency of PTH) . It causes the lowering of blood calcium level. This increases the excitability of nerves and muscles, caus

Pvcs during recovery, As the heart rapidly slows during recovery from exerc...

As the heart rapidly slows during recovery from exercise, PVCs commonly occur and usually have no clinical significance. This may be a time when metabolic adjustments in the heart

Causes of apomixis, Causes of Apomixis Apomictic species are generall...

Causes of Apomixis Apomictic species are generally hybrids or polyploids, as a consequence, there is irregular meiosis. Apomixis appears to be controlled by a set of genes. T

Availability of adp, Availability of ADP When  the ADP  levels increase...

Availability of ADP When  the ADP  levels increase due to hydrolysis ofATP  in various biosynthetic reactions,  the  rate of reaction to generate ATP  is accelerated and this i

Excreation, what is the role of liver for the metabolism of carbohydrates

what is the role of liver for the metabolism of carbohydrates

What is digestion, What is digestion? Digestion is the breaking down of...

What is digestion? Digestion is the breaking down of larger organic molecules obtained from the diet, e.g. carbohydrates, fats, proteins, into smaller ones, as glucose, fatty a

Hypertension, Define hypertension, discuss its causes and explain its effec...

Define hypertension, discuss its causes and explain its effects on the body. What role, if any, does medical imaging play in hypertensive patients? Explain and give examples.

What is the main cell organelle involved in cell digestion, What is the mai...

What is the main cell organelle involved in cell digestion? What are the properties of that organelle that enable it to do the task? The organelles responsible for intracellula

How parasitic infections are found, How Parasitic infections are found? ...

How Parasitic infections are found? Parasitic infections are found throughout the world. With increasing travel, immigration, use of immunosuppressive drugs and the spread of A

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