IGEM:IMPERIAL/2006/Protocols/mini prep of culture

Miniprep of cultures
Solution 1 Solution 2 Solution 3
 * Transfer 1.5ml of culture into eppendorf tubes.
 * Spin for 20seconds and remove supernatant.
 * Take out Solution I and III from the fridge with syringes. Solution II (if it is cold it precipitates) is on the bench. (use yellow pipette for syringes)
 * Add 100ul of solution I and resuspend the pellet by vortexing.
 * Add 200ul of solution II.
 * Add 150ul of solution III.
 * Mix them well by inverting the tubes. White precipitates appears.
 * Spin for 3min. In the meantime, label tubes.
 * Pour the supernatants in to the new labelled tubes, add 1ml of ethanol and mix.
 * Spin for another 3min and remove supernatant.
 * Spin to remove residual liquid and then resuspend in 50ul of MiliQ (1 sec. vortex. Don’t resuspend the pellet).
 * NB: If the culture was grown during the day (8hrs), then resuspend in 25 to obtain approximately the same concentration.
 * Spin for 1min.
 * Ready to digest.
 * 90g glucose
 * 10ml 0.5M EDTA
 * 6.25ml 2M Tris pH8
 * make upto 500ml
 * For 500 mls
 * 50ml 10%SDS
 * 20ml NaOH
 * 147g potassium acetate
 * 57.5ml glacial acetic acid
 * make upto 500ml