I am currently doing DNA dot blots using nitrocellulose, however, i need to increase DNA binding thus thought of using PVDF membrane. What method of activation should I be using on the PVDF. I find that DNA resuspended in Tris/NaOH doesn't get absorb into the PVDF membrane.
You will have to wet it with methanol as mentioned above. In addition, may be the the link below is interesting to you. Good Luck
http://www.pall.com/main/oem-materials-and-devices/literature-library-details.page?id=27294
Hi, I agree with the opinions above but I would suggest you to use Hybond +. In my dot blot experience I am using always Nylon + membrane and to fix the DNA to the membrane I treat the blot with NaOH as classical dot blot but is not fixing very well so I do additional step including baking 2h at 80 deg. which of course is not suitable for nitrocellulose. Good luck!
See this info published on Pall website it is everything you want to know about nucleic acids and PVDF blotting. All biomolecules stick to these membranes via hydrophobic interaction.
http://www.pall.com/main/oem-materials-and-devices/literature-library-details.page?id=27294
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