I am staining cells with dapi, which thought to bind A-T rich region of double helix. and we know that dapi fluorescence intensity increases when attached to DNA as compared to unbound state. DNA excited by UV at 358 nm with emission peaks at 461 nm. Also weak fluorescence can be detected for RNA binding which emission shifted to 500 nm.
Is it possible to measure the amount of emission by DNA and/or RNA separately. Particularly interested in RNAs emission (above 500nm). Thanks
What kind of sample and hardware are you using (Cells on a slide, homogenates, microscope, fluorimeter)?
Have you tried to use a fluorimeter and titrate your assay with DNAse treated-RNA extracts? DAPI-RNA signals might be very weak to be usable. Also, other compounds are known to bind to DAPI and emit fluorescence at 500+ nm.
I agree with the answer posted from Fabio.
Please start Google Scholar and not Google and insert your quesion .
you' ll find 21.600 hits. Please search and read these references. It is always worthwhile to search in Google Scholar prior posting a question.
see too:
http://www.tandfonline.com/doi/abs/10.3109/10520299509108199
JRG
You can measure the two different emission of same fluorophore by using filters of defined wavelength. For example in case of DAPI, you can use emission filters en-flanking 461 nm and 500 nm wavelength sequentially with same excitation wavelength of 358 nm. I have used Zeiss Confocal microscope where you have tools for this type of analysis.
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