User:Anthony Salvagno/Notebook/Research/2010/02/15/More Tethering with DNA Concentration

I am planning this for this afternoon because Andy is doing microtubule stuff and I want to get a rough poster plan set up.

What to do today
Last week I did some experiments that dealt with larger concentrations of DNA and got some pretty good results. Today I will do some more: I will do 1:20, 1:50, and 1:100 today. I'm hoping that that is enough to get valid information. Although it would be good to do it all because the concentrations I deal with for unzipping are significantly less than those from PCR (probably about 100x less concentrated).
 * Starting amount
 * 1:5 dilution - 2ul start in 8ul water
 * 1:10 - 1ul start in 9ul water
 * 1:20 - 0.5ul start in 9.5ul 1x Pop
 * 1:50 - 1ul start in 49ul 1x Pop
 * 1:100 - 1ul start in 99ul 1x Pop
 * 1:500 - 1ul 1:50 in 9ul 1x Pop
 * 1:1000 - 1ul 1:100 in 9ul 1x Pop

Setup
I still have 2 more sets of cleaned glass so I will use those for today. I will also dilute the beads 1:25 in 50ul and sonicate for clump removal. Also I need to make more BGB.

Making BGB
I want a large unfiltered supply from which I can filter and aliquot out. I think I will go with the smaller falcon tubes full of popping buffer and the appropriate amount of BGB. So that's 15ml capacity which gives me 0.075g BGB. Cool beans.
 * Goal: 5mg/ml BGB in Popping Buffer

To be continued...
Tomorrow