Difference between revisions of "Miniprep"

Revision as of 00:54, 19 September 2014

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:

@@ Line 1: / Line 1: @@
 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:
 [[File:AM-112113_1.jpg]]