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Abstract for AluQuant® Human DNA Quantitation System
The AluQuant® Human DNA Quantitation System uses two
incubation steps to measure the amount of human DNA. In the first incubation,
a coupled enzymatic reaction takes place. The first reaction is known as a
pyrophosphorylation reaction. This reaction is the reversal of the DNA
polymerization reaction, where a DNA polymerase, known as the READase™
Polymerase, catalyzes the addition of a pyrophosphate across the 3′-terminal
bond of double-stranded DNA. This addition results in the release of the
3′-terminal base from the DNA strand as a deoxynucleoside triphosphate
(dNTP). The terminal phosphate of the released dNTP is then transferred to
ADP to form ATP using the second enzyme in the reaction mixture, the
READase™ Kinase. In the second incubation, the amount of ATP produced
in the first reaction is quantitated by measuring the amount of light produced
from the luciferase activity in the reaction. This system uses DNA probes
that are specific to repetitive human elements allowing quantitation without
PCR amplification.
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