How much of the fluorescently labeled PCR probe (500bp) will be enough for the northern blot.?

08 November 2022 1 6K Report

I have been trying different concentrations (1259ng, 2250ng) in hybridization solution (50ml) but no signal. (I tried with control also where there was just the probe against probe).

Or is it something else other than the concentration, like something wrong with the label?

Really looking forward to some suggestions.

Thanks in advance.

Frederic Lepretre

Hi Saleha Arooj

first thing is are you sure of your probe, before quantity you should try quality. put some on agarose gel and see if there is something at the right size. if yes, you should try to see if it suits your target, by blasting the sequence in NCBI or UCSC.

if no issues, maybe your labeling protocol was wrong. every steps must be checked...

all the best

fred

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