Yesterday when taking data I was fighting with DNA issues. First there was too much and then it all but practically vanished. Then today, I had absolutely no problem obtaining a good coverage of DNA, but the NucS was practically nonexistant. I was using half the amount of buffer to combat this issue and twice the volume of the protein and yet I am still not really satisfied with the number of interactions I could see while filming.
I just started the analysis of the first video from today, so I will soon see if enough data points came out of it. It could be that the very few proteins I do see are actually the only ones interacting and that for some strange reason, BSA is being very robust today in preventing the non-specific interactions. I checked the concentration of the NucS on a slide all by itself and it really did seem fine so I do not think thats the problem.
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