Difference between revisions of "Miniprep"
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[[File:AM-112113_1.jpg]] | [[File:AM-112113_1.jpg]] | ||
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+ | * For alkaline lysis step leave for 5 mins | ||
= Colony miniprep = | = Colony miniprep = | ||
As above but pick a colony to do the kit miniprep process with rather than growing a liquid culture. | As above but pick a colony to do the kit miniprep process with rather than growing a liquid culture. |
Latest revision as of 19:00, 6 May 2015
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.