IGEM:Harvard/2006/Adaptamers/Notebook/2006-8-13

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8/13

Attracting streptavidin magnetic beads with the 8/9 setup doesn't seem to work well: attraction to the side makes it difficult to discern a difference between setups. Orienting the magnet vertically seems a little more promising since it is the strength of the magnet directly against gravity; hopefully the gradients of magnetic particles will differ based on differing properties of large aggregates (densities, magnetic bead: thrombin bead ratio, surface area (and friction) differ. The vertical setup yields very little pulling power even for .2 mL PCR tubes, so I cut a couple up to get the magnet in range. This experiment is questionable as I reused the refrigerated 8/10 experiment beads (incubating on a rocker for 30 minutes).

I also reran streptavidin plus the various types of thrombin we've used and BSA on a SDS-PAGE 12% Tris-Glycine gel for 1.5 hours. Results shown below.

Lane Condition
1Seeblue plus2 ladder
28 uL 1% BSA
340 pmol Haematologic Technonlogies human alpha thrombin
440 pmol Sigma Human thrombin
540 pmol Bovine Thrombin w/o BSA
690 pmol streptavidin

Image:cst813proteins.jpg

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