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11)add 250ul nuclear extract buffer NEB to pellet,stand on ice5', gently resuspend and transfer to dialysis tubing (cut©off14kD, no azide) 12)dialyze 4h x 2 x 2l NEB
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2. Add 250ul of Buffer S1 to the pellet to resuspend bacteria cells. 3. Add 250ul of Buffer S2, mix gently by inverting the tube 4-6 times until the solution becomes clear. The time should be no longer than 5 minutes. 4. Add 350ul of Buffer S3, mix gently by
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initials and date on each conical. Place in the -20 qC, TC lab. Brefeldin A We order BFA in 5mg vials (Sigma #B7651). Reconstitute the BFA with 1mL DMSO (5mg/ml) and then create 5 working solutions by aliquotting 200µl of the resuspended BFA into1.7mL