User:Brian P. Josey/Notebook/2009/12/09
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From the tethering yesterday and last week, it is clear that the beads we are using are sticking together and we need to separate them from each other to avoid clumping. Anthony is showing me how to do this. We spin it down at 6.6g for 5 minutes. We then remove the liquid from the centrifuge tube by pipetting it out. We then replace it with 950μL of the popping buffer, and repeat three to four times. Anthony wrote a page on the protocol in his notebook that goes into more detail. This serves to remove the streptavidin. We then used sonication to break the beads up and prevent them from clumping together. This presents the streptavidin from interfering with the tethering, and keeps the beads from sticking together.
I am also going through my notebook and trying to update it and clean it up. I will at least get through yesterday's page.