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FISH (Fluorescent in situ hybridization) What volumes are recommended for buffer


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#1 Amigo

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Posted 01 February 2011 - 04:13 PM

Hi everyone,

I wonder if anyone can give me some advise on what volumes to use for FISH buffers. My samples are small (lenght 1-2mm) and .. not sure how thin... very thin. Anyways, how much hybridization buffer to people usually use? just enough to cover the tissue? how much washing buffer?

Or I guess the more general question is, is the volume relevant? The protocol I was given says I need 5ng/ul of probes. So I will use that concentration but as I said I am not sure on how much volume to add. I should say I have never done FISH before.

Thank you in advance,

Amigo

#2 bob1

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Posted 02 February 2011 - 05:38 PM

Volume isn't too important other than it should be enough to cover the sample(s) well. It is better to use larger wash volumes though, as you are trying to remove the unbound probes etc.

#3 Amigo

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Posted 03 February 2011 - 12:38 PM

Hi Bob 1,

Thank you for your reply. Cheers,




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