BIOL 301 Lecture Notes - Immunoglobulin G, Cuvette, Reagent

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Add 200 ul of lysis buffer to the tube containing the yeast pellet and vortex until cells are completely resuspended in the buffer. Pour all the beads (200 ul) into the previous suspension. Vortex the tube for 1 minute at high. Place on ice for 1 minute. (always keep on ice from now on) Repeat steps 3 and 4, 7 times. Spin for 5 minutes at 2000 rpm at 4c. Transfer supernatant (200 ul) to a new tube. Add 6 ul rnase/dnase to the supernatant. Add 300 ul of extraction buffer and invert the tubes 4-5 times (avoid foam) Take tubes on ice to 4c fridge and clamp them in the rotator for 30 minutes. Place the labeled tubes in the refrigerated centrifuge. Spin at 4c for 10 minutes at 12000 rpm. Transfer the supernatant (400 ul) to a micro tube marked total and place on ice.

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