I measured the DNA concentration using Nanodrop 2000 spectrophotometer and ROCHE High Pure PCR Template Preparation Kit. In the Nanodrop result, the screen shows contaminants in the DNA sample, as represented in the attached screen capture.
Did you use your TE buffer/DNA elution buffer to blank the spec?
Looks to me like you either: don't have DNA, didn't properly blank the spec, or the pedestal is dirty.
Also, the low A260/A280 ratio means you have a lot of contaminants present.
Clean the pedestal, blank the spec, and try again.
Good luck!
Did you add absolute ethanol to all wash buffers before using?
I think you can keep the pedestal clean by using nuclease free water twice before using it. Also, you can use nuclease free water as blank.
Exactly we had prepared wash buffers with absolute ethanol 100 % according to the manufacturer protocol. Ethanol doesn't have a significant effect on the absorbance spectrum of DNA when measured in TE pH 8.0.
Thank you for your kind response dear Asif.
Hi,
Did you perform the quantitation with other samples? Perhaps the problem may be in the calibration of the equipment.
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