Hi, I am Ashila. I'm working with DNA macroarray right now but still got no dots on the membrane. I'm using nylon membrane from Roche and mostly Roche products for my exp. I have troubleshoot some steps but still not sure which part is the problem.
Steps:
1. Prepare plate - 20 µl of spotting buffer + 20 µl of oligo 30 pmol
2. Nylon membrane printing using 96 pin replicator, dry 1 h
3. Crosslink using UVP ClL-1000 240mJ
4. Incubate 60C 1h
5. Wash - 0.5% SDS 60C 1h, rinse 100mM Tris at room tem 5 min
Next is just followed protocol by DIG Easy Hyb (Roche)
I got no result even for my +ve control. Need some suggestion here. TQVM