I have performed Dot Blot assay, but the background was too high.
For washing the membrane PBS-Tween was used. Can anybody suggest other washing buffer?
Hi Agnes
I should advice you to use the "Maniatis" protocol.
Standard Washing:
- wash 2x for 15 min in 50 ml of 2x SSC, 0.1 % SDS at room
temperature
- wash 2x for 15 min in 50 ml of 0.5x SSC, 0.1 % SDS at 55 ̊C
(prewarmed)
or
for higher stringency:
wash 2x for 15 min in 50 ml of 0.1x SSC, 0.1 % SDS at 60 ̊C
Hope it helps
Best
Paco
Thank you Francisco!
I have never performed FISH experiment on Arabidopsis.. I need a detailed protocol.
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