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

Why dairy or milk product are important for human body, Why Dairy or milk p...

Why Dairy or milk product are important for human body? This first food of mammals is rich in body-building proteins and bone-forming calcium, besides being the only source of

Explain some ethological activities, Explain Some Ethological Activities ...

Explain Some Ethological Activities Comparative studies of behaviour or activities are also used in systematic for identification of organisms. The comparative study of the beh

Wetlands - lentic ecosystems, Wetlands - Lentic Ecosystems Wetlands a...

Wetlands - Lentic Ecosystems Wetlands are permanently or periodically water covered areas. They can be defined as submerged or saturated lands either artificially or naturall

Operculum - seed appendages, Operculum - Seed Appendages The term oper...

Operculum - Seed Appendages The term operculum is applied to a plug-like structure formed in the micropylar portion of the seed by proliferation of cells at the tip of the inn

Considerations for congestive cardiac failure, Q. Considerations for conges...

Q. Considerations for congestive cardiac failure? - Subjects with congestive cardiac failure often tolerate small frequent meals better than larger infrequent meals as these a

Membrane that selects based on pore size, A transport process in which phys...

A transport process in which physical (hydrostatic) pressure drives part of a fluid through a membrane that selects based on pore size

Heart output, HEAR T OUTPUT The amount of blood pumped by heart per...

HEAR T OUTPUT The amount of blood pumped by heart per minute. Heart beat is 72. Pump out blood is 70 ml. This 72 × 70 = 5040 ml. ( 5 litres) blood is pumped in a minute.

Fate of blastopore, Fate of Blastopore Cleavage results in the formati...

Fate of Blastopore Cleavage results in the formation of a ball of cells called morula (resembling mulberry hence the name). A space appears in the morula changing it to a holl

Screening dna libraries, Genomic  libraries  are  screened  by  hybridizati...

Genomic  libraries  are  screened  by  hybridization with  a  DNA probe which is complementary  to component of the nucleotide  sequence  of the desired gene. The problem should be

Trophic structure-structure of community, Trophic structure Organisms i...

Trophic structure Organisms in a community are closely interrelated with each other through feeding relationships. Another aspect which is quite obvious in a community is th

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