Making tethers with Pranav & Priyanka * New BGB Weighed out in a boat 196 mg of Blotting Grade Blotter Added 19.6 mL of 1x Popping buffer Sterile filtered into 50 mL falcon tube. This made "2x BGB," approx 10 mg / mL BGB, assuming not a lot of protein stuck to the filter Made 1x BGB by adding 750 ul of popping buffer to 750 ul of 2x BGB. Mixed by aspiration * New Beads Took 100 ul of stock beads CP01F Lot 8892 Actual 196 mg, add 19 ml 1x Pop Diluted 5 ul + 45 ul 1x popping buffer. Sonicated for about 30 to 60 seconds, significant splashing. Quick spun to get off sides. * New Antidig Diluted fresh aliquot of antidig with 180 ul of cold 1x PBS * Clean glass Cleaned using just water and scrubbing, followed by water rinse about 6 times. My intuition is that this did not clean the glass well at all. The tape would not stick to the glass, way worse than I've ever seen it. Subsequently, both samples leaked. Sample may or may not Sample Antidig for about 4 minutes 5 s.v. BGB, 3x. Top sample... - Steve Koch