Hi friends, let me explain my query in detail.
By analysing a ladder standard on Micro spectrometer I am getting answer 505ng/ul, which is quite correct, the standard is of 500ng/ul. Similarly, I analysed extracted sample of a DNA the reading showed, 3300ng/ul.
Same samples were analysed on gel electrophoresis. Considering the ladder concentration and its band intensity obtained on gel electrophoresis, comparing it with the sample's intensity, results is 1810ng/ul concentration, which is almost half of the value obtained by micro spectrometer .
I am not able to understand how two techniques are giving two different results for the same sample, viz : 3300 and 1810.
Please help. Thanks in advance.
Spectrofotometry measures everything that absorbs at 260 nm (do you see the spectrum or only the absorbance value?), including nucleotides and/or RNA.
When you've run the electrophoresis, have you seen single band?
Hi there,
As Tomas mentioned, Spectrum (260/280 and 260/230 ratio are very informative) and gel picture would help to have a better analysis. If DNA has been silica gel purified then it's not nucleotide contamination, RNAse treatment should eliminate RNA contamination and in case of PCR amplified DNA, gel purification or column purification should eliminate residual primers and secondary products.
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