I am crosslinking DNA sample on hybond N+ membranes. After spotting I let my mebrane dry for 30 min to 1 hour. Then comes the crosslinking step. Should I have the mebrane soaked in TBST buffer and then perform UV crosslinking or I can use the dry membrane itself for linking the DNA?
Thank you.
After soaking it in TBST, air dry it and go for crosslinking.
Agnishwar Girigoswami
please let me know if you mean following and also suggest few queries:
2. dry for 1 hours as I do
3. soak in TBST for how long??
4. air dry again (for how long)
5. UV crossslink (I do 1 autolink)
Thank you
Agnishwar Girigoswami 15 min is standard for what: drying post TBST or soaking in TBST?
Thank you
Here is what I do now and it is working: I add samples onto membrane using dot blot apparatus from Bio-rad. I put the membrane immediately onto water soaked blotting paper in a plastic tray. I perform UV crosslink (1 autocrosslink) and remove the blot from the tray and put it in the buffer (TBST) for 2 min and then transfer the blot into blocking solution. I do not dry the membrane throughout the process and it works all okay.
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