Gradient preparation

Place the micro leveler on the rotator and adjust until the bubble is centered. Press the level and ensure the plate is absolutely horizontal.

Place the ultracentrifuge tube in the marker block and mark at the 50 percent position for the short cap.

Draw the 10 percent sucrose solution into a 10 ml syringe with wide gauge cannula and add it slowly to the bottom of the tube until reaching the 50 percent mark, ensuring no bubbles form.

Carefully add the 50 percent sucrose solution to the bottom of the tube, layering it under the 10 percent solution until the lighter solution rises nearly to the top, leaving approximately 1 mm space.

Insert the short cap into the tube ensuring a small amount of sucrose comes out to eliminate any remaining air bubbles at the top.

Select the appropriate program that specifies rotation time and angle, then press run to tilt and rotate the tube, creating the gradient.

Allow the rotator to complete the rotation cycle and return to vertical position. Store the completed 10 to 50 percent sucrose gradient at 4 degrees Celsius.