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

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

Contents

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:

  • 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

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).

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

  • Goal: 5mg/ml BGB in Popping Buffer

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.

Note to self - I need more Popping Buffer

To be continued...

Tomorrow

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