I was wondering whether it is nescessary to crosslink DNA to a nylon membrane (for dot blot assay preparation), or is air-drying the DNA onto the membrane sufficient for strong binding prior to probe interrogation?
Hi there,
DNA interacts with nylon membrane through strong hydrophobic interactions. So it is not compulsory to stabilize the interaction with crosslinking. Nevertheless and depending on the condition used for hybridization, crosslinking might improve the signal.
Hi Adam,
From my experience I would recommend crosslinking by uv radiation. If you want to perform reverse dot hybridization you can "adjust" the angle that your probe needs to bind to the membrane by different exposure times. The angle is important for the quality of your hybridization.
Best regards
Dirk
Can anyone tell me the protocol for UV cross linking of DNA aptamer on NC membrane
Following Tammineni's question, what range (nm) of UV can be used for crosslinking DNA onto nylon membrane. Thanks
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