User:Anthony Salvagno/Notebook/Research/2009/10/15/Making Tethers

Supplies

 * I made 1x popping buffer
 * I made k-casein in 1x pop (5mg/ml) hopefully this is a good blocker
 * Need a-dig
 * DNA
 * beads

A-dig
For this there are 20ul aliquots and I need 1:10 of that so add 180ul 1x Pop

DNA
I can try and tether both strectching DNA 4.4 kb and some unzipping stuff. I should dilute about 1:10 fold for both so that I can see if I got anything. I think we went more dilute, but I want tethers.

Beads
I need to dilute beads (amount TBD after looking through notebook), then sonicate to remove clumps. Following this protocol: User:Anthony Salvagno/Notebook/Research/2009/08/06/Bead Washing

OK Beads are now made. I don't think sonicating worked well.

Tethering
Ok, now everything is ready. It is time to tether.


 * 1) Flow a-dig
 * 2) *allow to sit 5min
 * 3) flow BGB
 * 4) *allow to sit 2min
 * 5) flow DNA
 * 6) *flowing 4.4kb stretch DNA and TD(1) unzipping DNA
 * 7) *allow to sit 5min
 * 8) wash with Pop
 * 9) flow Spheres
 * 10) *allow to sit 7min
 * 11) wash with pop
 * 12) Seal with nail polish