Hi.
I agree with Parisa. I also don't use frosted slides for comet assay. I use normal slides (I specifically use 3 window diagnostic slides).
Also, preferences vary (some coat with NMPA first and LMPA second), but I coat the slides with 0.75% LMPA twice as thus;
I coat each window with 15ul of 0.75% LMP Agarose first. Dry these in the incubator at 37 degrees for 30 minutes. Thereafter, i coat again with 15ul of 0.75% LMPA and allow to dry at least for 24hours before use. I think the longer you keep them the better. Perhaps prepare your slides between 2-5 days before your assay and leave them in the incubator.
Note: if you are using the conventional slides (not the 3 window disgnostic slides), you'll require a larger volume of the LMPA to coat. Perhaps about 50-75ul as opposed to the 15 ul twice that I used.
All the best.
Dear Neethu,
Frozen slides are good for better attachment of NMA and you will get a uniform layer.
as suggested by Finosh GT, frosted side have rough surface, which help in better attachment of the NMA layer while slide preparation.
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