Comet assay - AGAR not sticking - (Aug/16/2005 )

I have been doing a number of comet assay's on fish blood and have found that the AGAR quite often falls off the slide during lysis or in many of the washing stages. I was hoping people may have some hints as to how to avoid this problem.

I have heard that heating the slides slightly before adding the agar/cell suspension may help?

i guess u are doing comet assay on slides

u could lay the first layer of gel and just flame it briefly. that should help the gel stick.

ya i also clean the slide with 70% ethanol v´before casting i guess u do the same

hope this helps

prajwal

It could also be that you electrophoresis buffer was warm, keep it in the fridge after preparation. Also ensure that the LMPagarose has set before you start processing the samples. When you think the gel has set, ensure that there is a 0.5mm dried ring due to agarose disc retraction at the edge of the comet slide area.

cold slides at 4 degree after adding the agarose