Miniprep

Anachem miniprep protocol (AM-112113)

Prior to miniprep, overnight incubation:

• Prepare 100ml of LB broth, using 2.5g of dry mixture per 100ml of water (this is also written on the side of the LB pack)
• Vortex, cover, add autoclave tape, and autoclave for 15 minutes.
• Let the autoclave cool. Verify that the tape has changed colour (the green-striped tape goes dark but not completely black)
• Ensure the mixture is cool (<40 degrees?)
• Transfer to the linear flow area. Ensure the area is sterile using 70% ethanol solution. Wear gloves and disinfect them. Do not add anything to the linear flow area unless it is sterile.
• Add antibiotic. For example, ampicilin stock is in the freezer. The stock solution is 100ul/ml. Add it to the broth at a ratio of 1ul per 1ml, i.e. add 100ul.
• Innoculate with your bacteria.
• Cover, label, and incubate at 37C.

Miniprep:

• For alkaline lysis step leave for 5 mins

Colony miniprep

As above but pick a colony to do the kit miniprep process with rather than growing a liquid culture.