Team:Stanford/NotebookPage/30 June 2010

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(Difference between revisions)
(New page: ==Laura's Lab Notebook:== Re-try failed transformation (failed 2X for Alex and Chris) Protocol followed: #get competent cells from -80oC freezer #set H2O bath to 42oC (already done) #add 5...)
(→Laura's Lab Notebook:)
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#set H2O bath to 42oC (already done)
#set H2O bath to 42oC (already done)
#add 50 uL cells to a 1.5 mL tube (keep tubes on ice)
#add 50 uL cells to a 1.5 mL tube (keep tubes on ice)
-
#add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL this time)
+
#add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL I0500 this time)
#on ice 30 min
#on ice 30 min
#24oC H2O bath 20 sec.
#24oC H2O bath 20 sec.

Revision as of 19:10, 19 July 2010

Laura's Lab Notebook:

Re-try failed transformation (failed 2X for Alex and Chris) Protocol followed:

  1. get competent cells from -80oC freezer
  2. set H2O bath to 42oC (already done)
  3. add 50 uL cells to a 1.5 mL tube (keep tubes on ice)
  4. add 50 ng or 2.5 uL circular DNA to E. Coli (variation: used 5.0 uL I0500 this time)
  5. on ice 30 min
  6. 24oC H2O bath 20 sec.
  7. on ice 15 min
  8. add 1 mL SOC, 2 hours at 37oC (rotating)
  9. plate100uL on appropriate plate (kanamycin this time), incubate 12-16 hours before picking colonies
    1. variations: 200 uL plated this time, also plated on kan/X-gal/IPTG plate (plasmid inducible copy # by X-gal)
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