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Generating Probe for Northern Blotting

Generating Probe for Northern Blotting - Biology Forum

Generating Probe for Northern Blotting - Biology Forums. Ask questions and discuss the study of Biology. If you have Biology questions from your homework ask them here!


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Old 06-27-2011, 06:09 PM
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Smile Generating Probe for Northern Blotting



I have the sequence information for a piece of DNA I would like to use as a probe in Northern blotting. Is designing primers and then performing PCR the best way to generate this probe?
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Old 06-28-2011, 06:02 PM
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Default Re: Generating Probe for Northern Blotting

I think the best way to make a probe for Northerns is to make antisense RNA probes. These apparently give the strongest signals and are of course strand specific.

In our lab, we Klenow label DNA fragments using random primers and this works OK, although the sensitivity is not that fantastic.

I would think that PCR using labelled dNTPs would also give as good if not better results than Klenow labelling.
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