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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.
How have prokaryotic cells given origin to aerobic eukaryotic cells and to photosynthetic aerobic eukaryotic cells? As per to the most accepted hypothesis aerobic eukaryotic ce
How are ecological interactions classified? Ecological interactions are divided as intraspecific or interspecific interactions and as harmonious or inharmonious interactions.
In this unit you have learnt that heart failure is one of the most common conditions seen and results in a high mortality. It can result from failure of the heart muscle as with co
Name two stimulant drugs and state the undesirable side-effects of each. Amphetamine and cocaine are stimulant drugs. The side-effects of amphetamines are high blood pressure;
Define Absorption, transport and Excretion of phosphorus? As you have seen that food contains both organic and inorganic phosphorus, but most of it is absorbed in its inorganic
wolves are predators for hares the wolf population increases what happenes next?
How do organisms adjust to changes in temperature? Some of the most ordinary way for an organism to adjust to changes in body temperature is by perspiration or panting.
what are the two main pollutants that contribute to acidic rain?
Zygote - Embryogenesis The fertilized egg or zygote is situated at the micropylar end/pole of the embryo sac, its basal (micropylar) end is attached to the embryo sac wall and
Explain Functions of Ascorbic Acid? Vitamin C is easily oxidized, and the majority of its functions in vivo rely on this property. It plays a key role in the body's synthesis o
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