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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.
Nuclear endosperm in angiosperm.
Define some Usual Doubts of Children related to Food? Before embarking yourself to go through this unit, here is an activity for you to perform, Talk to 2-3 school' boys and gi
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Database Search: Once an open reading frame or the partial amino acid sequence has been pre-determined, the investigator compares sequence with others in the databases using a com
Define working of Body surface in excretion of zinc in human? Loss of zinc occurs due to the exfoliation of skin and sweating (0.7 -1.0 mg/day). Another route of zinc loss is h
What are likely to be (a) the coldest, (b) the warmest parts of the body? The extremities of the body (hands and fingers, feet and toes, ears and nose) are likely to be the col
State two ways in which sulphur dioxide emissions from coal-fired generating stations could be reduced. Sulphur dioxide emissions can be reduced by fitting desulphurization pl
what is mean by autotrophic ?
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differentiate between scanning electron microscope and a transmission electron microscope in terms of their functions
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