Difference between revisions of "Miniprep"

From London Hackspace Wiki
Jump to navigation Jump to search
 
Line 15: Line 15:
  
 
[[File:AM-112113_1.jpg]]
 
[[File:AM-112113_1.jpg]]
 +
 +
* 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:

AM-112113 1.jpg

  • 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.