User:Daniel Goodman/Notebook/Cluzel/2010/03/31: Difference between revisions

Latest revision as of 18:10, 31 March 2010

Cluzel Lab Notebook
Daniel Goodman

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Grow 250 ml flask, one colony, 50 ml LB at 37 deg for 4 hours (OD must be 0.3-0.4) in shaker (check at ~7pm) - OD was 0.83

Spin down 20 mls in 2 50 ml epis at 4 degrees in centrifuge
- 3200 RCF (4000 RPM) 10 minutes 4 degrees
- pour of supernatant, add 40 mls 4 deg filtered water each (maybe add 5 first, vortex, add 35 more)
  - make sure pellet is dissolved
- repeat above 2x
resuspend pellets in 10% glycerol (5 mls each)
spin, remove glycerol supernatant
resuspend in 0.5 ml 10% glycerol each, combine tubes
make 20x 50 ul aliquots, put in -80 freezer

Put silicon chips in acetone (under fume hood) in glass flask for 5 minutes, heated slightly
Made 3ml 5% agarose, heat to 80 deg
Using stationary phase GFP cells, dilute a few colonies in DI water to OD 0.05.

@@ Line 1: / Line 1: @@
 {{User:Daniel_Goodman/Notebook/Cluzel/Template:Entry_Base}}
-==Entry title==
-* Insert content here...
-Protocol for Competent Cells
+==Protocol for Competent Cells==
+*Grow 250 ml flask, one colony, 50 ml LB at 37 deg for 4 hours (OD must be 0.3-0.4) in shaker (check at ~7pm) - ''OD was 0.83''
-*Grow 250 ml flask, one colony, 50 ml LB at 37 deg for 4 hours (OD must be 0.3-0.4) in shaker (check at 7pm)
 *Spin down 20 mls in 2 50 ml epis at 4 degrees in centrifuge
 **3200 RCF (4000 RPM) 10 minutes 4 degrees
@@ Line 16: / Line 15: @@
 * make 20x 50 ul aliquots, put in -80 freezer
+==Agarose Chip Prep==
+* Put silicon chips in acetone (under fume hood) in glass flask for 5 minutes, heated slightly
+* Made 3ml 5% agarose, heat to 80 deg
+* Using stationary phase GFP cells, dilute a few colonies in DI water to OD 0.05.
 |}
 __NOTOC__