Team:Alberta NINT/Protocols

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Lab Protocols

Coming Soon!

Overnights

Inoculating LB culture tubes:

   Add 5ul Amp50 or Amp100 to 200ml LB broth in tube.
   Using sterile inoculating loop, touch an isolated colony.
   Swirl loop through LB + amp broth.
   Incubate overnight at 37 C.

Minipreps QIAprep Spin Miniprep Protocol

Isolating DNA from LB culture tubes:

   Add 2X 750ul broth from overnights into a 1.5ml tube.  Centrifuge for 1 min to pellet and aspirate liquid.
   Add 250ul Buffer P1 and resuspend pelleted cells.
   Add 250ul Buffer P2 and invert tube 4-6 times.
   Add 350ul Buffer N3 and invert tube immediately 4-6 times.
   Centrifuge for 10 min.
   Apply supernatant to QIAprep spin column.
   Centrifuge for 1 min and discard flow through.
   Add 500ul Buffer PB, centrifuge for 1 min and discard flow through (only if high nuclease activity).
   Add 750ul Buffer PE, centrifuge for 1 min and discard flow through.
   Centrifuge for an additional 1 min.
   Place spin column in a clean 1.5ml tube.  Add 50ul Buffer EB, let stand for 1 min and centrifuge for 1 min.