I use Glycoblue to dye my RNA pellets deriving from cytosolic or nuclear RNA (concentrations ranging from 150 to 600 ng/uL) and everything is pretty fine with the absorbance ratios from Nanodrop.
However, when I do the same on RNA extracted from heavy polysomes (concentrations ranging from 65 to 170 ng/uL) I get very low 260/230 ratios (
I assume that A230 is from ribosomal proteins.
You have to realise that Nanodrop is useless for complex samples like polysomes (i.e., mixture of RNA and proteins).
Is there a more accurate fluorimeter available to you? The Qubit fluorimeter is much more accurate than the nanodrop since it's measures emission of a specific dye (binding specifically to protein, or RNA, or DNA), rather than just relying a absorbance at a specific wavelength.
Hi everybody,
sorry for the very late reply and thank you for your kind and helpful answers.
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