2 replies to this topic

[steffi333]

Hi all,

I'm looking to create a FISH probe from a PCR product - and from what I've read so far it seems I can accomplish this with digoxigenin or biotin labelling.

As far as I am aware, you can order primers with a DIG or biotin modification. Does anyone have any experience using DIG or biotin in PCR? Do you need to have special conditions or buffers etc?  

Does anybody have an opinion of which is better with regard to DIG or biotin? Or have any other suggestions? From what I've read DIG sounds as if it would be more specific from an antibody detection point of view, but in my experience that usually means you'll get charged more for it.

Thanks!

[pDNA]

the conditions for the PCR DIG labeling precedure can be found here!

Never used DIG-labled probes for FISH ...so i cant comment on the issue if DIG or Biotin is better.

Regards,
p

[phage434]

In general, the background with Dig is lower than with biotin labels, especially in a situation like FISH, where there will inevitably be biotin in the sample.