User:Anthony Salvagno/Notebook/Research/2010/06/25/Tethering in 3 different Environments

Quick Navigation: Pages by Category | List of All Pages | Protocols

Big beads are a bitch, and I'm going to make them mine!

The Quest

I need to get tethering to work. Period. So I will try tethering in 3 different ways:

1. Tethering with the beads in just Popping Buffer (not BGB)
2. Tethering with BGB beads at 37C
3. Tethering with BGB beads at 4C

This should be fun. Each sample will have a no DNA sample to keep tabs on.

The Prep

1. Wash small amount of beads (4k*g for 5 min), remove supernatant, resuspend in 50ul 1x Pop
2. Dilute anti-dig 1:100 in PBS
3. Dilute TpBR 1:100 in 1x Pop
4. Make sample chambers
5. Tether