User:Anthony Salvagno/Notebook/Research/2010/11/19/Tethering and PCR Results Part 2

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So yesterday I couldn't get definitive results if the PCR worked (both tries, the DNA stayed in the well) so I did PCR cleanup yesterday and am giving it one last try. On top of that I will try tethering with old DNA since I remade the Popping Buffer to better suit the needs of the experiment.

Update: The results of the gel show that the PCR was a failure. I don't know what could have happened, but I'll have to try again at some other point.

Tethering

I'm doing one sample of 1.1kb tethers and one sample of 4.4kb tethers (both stretching DNA). I had to reorder syringe filters so that I could filter my BGB, so in the meantime I used dephosphorylated alpha casein.

It turns out that I wasn't able to look at the samples until about 3 hours after I made it. There weren't any tethers after looking for a little bit, but there were some other interesting notes:

  • Thee beads weren't as clumpy as they have been the past week or so. I can attribute this to either the popping buffer being correct (the d-a casein was made over the summer when I had good popping buffer), or the casein working well. It should be noted that the beads were diluted 1:20 in the BGB I made last week from the improper Popping Buffer.
  • Also the beads were stuck. This could be because they had been sitting for so long. It could also be because the beads were tethers at some point and then got stuck.

On Monday I will do a sample of beads in new BGB and a sample of beads in d-a casein. I would like to figure out what the reason for the less clumps are.

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