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Estimation of DNA by diphenylamine
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Transcript
What is DPA ?..
Estimation of DNA by diphenylamine
T.A. Alaa Alahmadi
Practical...
Remember that ...
In DNA only the deoxyribose of the purine nucleotides reacts so that the value obtained represents half of the total deoxyribose present.
Principle...
1) Boiling of DNA under acidic condition
Causes depurination of the DNA followed by dehydration of deoxyribose sugar into a highly reactive hydroxylevulinylaldehyde.
The hydroxylevulinylaldehyde, under acidic conditions, reacts with diphenylamine to produce a blue-coloured complex that absorbs at 595 nm.
2) Treat DNA with DPA
unk.
Calculate conc. unknown (mg/ml- mg/dl- μg/ml & μg%), then the result multiple by 2.
Result
Diphenylamine is an organic compound
A diphenylamine indicator is often used for the detection of DNA.
DPA assay is quantitative test that used in biochemistry.
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