User:Anthony Salvagno/Notebook/Research/2010/10/19/Tethering To-Do

With the tweezer ready for tweezing, calibration under-way, and some last minute mechanical features being added it's time to get tethering, stretching, and unzipping going again. I need to get my head together to get it working at least as efficiently as it was before BPS last year. Yesterday I did some dabbling and it wasn't very good. Things need to be better.

To-Do List

 * make new BGB
 * I have a 50ml tube of it ready to go. It just needs to be filtered. I made this last June/July I think. Is it ok to go?
 * make new POP
 * I definitely need new pop and I think I need to remake one chemical in the mix.
 * prepare more anti-dig
 * Get new beads
 * small beads, the big beads are fine except they don't tether well
 * try cleaning with Windex
 * may be something magical in the windex that gave Koch great tethering. This is literally the only thing I've yet to do from Koch's glory days.

Tethering Thoughts

 * Right now I'm using 1:100 concentration for the 1.1kb tethers I think this puts it somewhere in the 10-100pM range
 * I need to mix my DNA tubes up. They've been sitting in the lab top coolers for some time now.
 * For DNA I have: TpBR (tetherable pBR fragments), Tyeast (I'm calling it that for now, it's some kind of shotgun clone), 1.1kb dsDNA tethers, 4.4kb dsDNA tethers (11pM I believe)
 * Since I'm starting with the dsDNA tethers (for stretching) should I use my frozen stock or will mixing it up do the trick? I guess I will try mixing first (gently of course) so I don't have to throw anything away.