User:Anthony Salvagno/Notebook/Research/2010/06/15/Tethering and Stretching Continued
Quick Navigation: Pages by Category | List of All Pages | Protocols
It seemed like yesterday we had a very major breakthrough. It seems there is a good chance that we overstretched DNA and I would like to try and continue that. It was also promising that we got tethers in the first place. So I have some things to follow up on.
Tethering
Ok so we got tethers, but they all seemed to break too easily. My theory is that the beads are barely getting close enough to attach to the DNA and so we are getting tethering events but the bond lifetime isn't optimized like it would normally be. I will try to tether with more NaCl in solution and giving longer bonding times. I will also do a No DNA sample. Here are my sample setups keeping everything else the same:
- No DNA 400mM NaCl
- 400mM NaCl + Beads no BGB
- 4M NaCl + Beads no BGB
- 50mM NaCl + Beads in BGB
- Beads in BGB (no added salt) plus 30min incubation time
I know this is 5 samples, but the last one will be it's own sample. I had to autoclave new tips and make new anti-dig.
Stretching DNA
Yea, we can overstretch DNA...