User talk:Brian P. Josey/Notebook/2009/12/09
Steve Koch 03:08, 10 December 2009 (EST): 66xg sounds unlikely? Also, sonication will be an important step. Actually this is my understanding: Sonication is useful for separating clumpy beads. Washing by cenrifugation/resuspension (which you are describing) is good for removing free streptavidin. Free streptavidin doesn't cause clumping, but we hypothesize that it will dramatically reduce tethering efficiency. Our reasoning is that free streptavidin will diffuse much more quickly than streptavidin-coated microspheres. Thus, a small concentration of free streptavidin could quench (bind to) all of the surface-tethered DNA, and ruin the whole assay.
Brian P. Josey 17:29, 10 December 2009 (EST) I couldn't remember if he had said 66x or 6.6x times so I wrote it down as 66x, I'll have to fix it. We did do sonication over in Evans' lab, so I'll add that step into my page.