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

Which functional group cross-linked in protein, Which functional group (sid...

Which functional group (side-chain) can be "cross-linked" in protein? a. sulfhydryl b. amino c. Phosphate d. Hydroxyl e. Carboxyl

Zoology, give detail account of modes of locomotion in protozoa and describ...

give detail account of modes of locomotion in protozoa and describe various type of asexual reproduction in protozoa

Would you expect the muscle fibers of the tongue, Would you expect the musc...

Would you expect the muscle fibers of the tongue to be striated or smooth? What about the muscle of the diaphragm/ Explain your answer.

Situation in which the mistakes holds in dna replication, Q. Mistakes may h...

Q. Mistakes may happen during each copying process. The same is true for the DNA replication. Are there correction systems in cells that try to mend those mistakes? Under which sit

Explain taxonomists as synthesisers, Explain Taxonomists as Synthesisers? ...

Explain Taxonomists as Synthesisers? Your study of the earlier units of taxonomy, in particular its history must have made you aware that the field of taxonomy has changed cons

Define the term - stimulus, Define the term - Stimulus Stimulus and res...

Define the term - Stimulus Stimulus and response characteristics of the tests themselves, as well as of the test instructions, become exceedingly important considerations. In g

Genetics, What is Mega sporogenesis

What is Mega sporogenesis

Define effect on oestrogens and progesterone in pregnancy, Define effect on...

Define effect on Oestrogens and progesterone in pregnancy? Oestrogens and progesterone are secreted by the placenta from 8-10 weeks of gestation. Progesterone increases through

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