User:Anthony Salvagno/Notebook/Research/2010/11/17/Tethering and PCR

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Today I'm tethering to see if tethering is working and I'm going to run a PCR of the 1.1kb stretching reaction.

Tethering

I'm testing to see if tethering is working in 2 ways. First I'm using different concentrations to see if beads are being tethered at all. In each case, the amount of beads should increase with increasing concentration (both stuck and tethered). So if I see more and more stuck beads I know something good is going on. Second I'm also testing to see if fresh from frozen DNA works better than DNA sitting at 4C. I'm hoping that I won't notice a difference, but I'm aiming to see if there are tethers with the frozen stock and no tethers with the 4C stuff. Here is my setup:

  1. 1.1kb at 1:100 concentration (I don't remember in molarity)
  2. 1.1kb at 1:1 concentration
  3. 4.4kb @ 20nM
  4. 4.4kb @ 2nM
  5. 4.4kb @ 200pM
  6. 4.4kb @ 20pM

If anything looks nice we will come back (or Pranav will) and try to do some stretching. In the meantime I will make new 1.1kb stretching DNA since I have only really old stuff and I like new stuff.

PCR

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