IGEM:IMPERIAL/2006/LabCalendar/2006-7-11

Miniprep- Prepare Plasmid DNA for electrophoresis
Follow miniprep protocol to prepare plasmid DNA samples for gel electrophoresis. Protocol for bacterial preparation for digestion:
 * Transfer 1.5 mL of culture into eppendorf
 * Spin for 20 sec and remove supernatant
 * Add 100 uL of solution 1 and vortex to resuspend
 * Add 200 uL of solution 2
 * Add 150 uL of solution 3
 * Mix by inverting, white ppt appears
 * Spin for 3 minutes
 * Pour supernatant in new labeled tube, add 1 mL of ethanol and mix
 * Spin for another 3 minutes and remove supernatant
 * Spin to remove resitudal liquid and then resuspend in 50 uL of MiliQ (ultrapure water) (1 sec on vortex, don't resuspend pellet)
 * Spin for 1 minute
 * Read to digest!

Make-up of solutions required for digest preparation
Solution 1
 * 90 g glucose
 * 10 mL 0.5M EDTA
 * 6.25 mL 2M Tris pH 8
 * Make up to 500 mL

Solution 2
 * 50 mL 10% SDS
 * 20 mL 5M NaOH
 * Make up to 500 mL

Solution 3
 * 147 g Potassium acetate
 * 57.5 mL glacial acetic acid
 * Make up to 500 mL