Haynes Lab:Notebook/Engineering PC-TFs/2013/02/05

(Difference between revisions)

Revision as of 16:01, 9 February 2013

Engineering PC-TFs

Bacterial transformation

Added total volume (10.0 μL) to 50 μL chemically competent cells (e.g., BL21) in a 2.0 mL tube.
Incubated on ice for 2 min., heat shock at 42°C for exactly 45 sec., and placed on ice.
Added 800 μL sterile SOC medium.
Grow with shaking at 37°C for 30 min.(Taped tubes to the shaker rack.)
Pelleted the cells at top speed in a microcentrifuge for 3 min. at room temperature.
Discarded the supernatant. Resuspended the cells in 100 μL LB + antibiotic.
Plate cells on pre-warmed LB agar + antibiotic. (I plated 100 μL of the cells. Then put beads on plates and shook.) Protocol didn't specify.) Grow overnight at 37°C.
Check results tomorrow.

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-==Summary==
+==2/5/13==
-*
+<font size=3>'''Bacterial transformation'''</font>
+* Added total volume (10.0 μL) to 50 μL chemically competent cells  (e.g., BL21) in a 2.0 mL tube.
+* Incubated on ice for 2 min., heat shock at 42°C for exactly 45 sec., and placed on ice.
+* Added 800 μL sterile SOC medium.
+* Grow with shaking at 37°C for 30 min.(''Taped tubes to the shaker rack.'')
+* Pelleted the cells at top speed in a microcentrifuge for 3 min. at room temperature.
+* Discarded the supernatant. Resuspended the cells in 100 μL LB + antibiotic.
+* Plate cells on pre-warmed LB agar + antibiotic. (''I plated 100 μL of the cells. Then put beads on plates and shook.) Protocol didn't specify.'')  Grow overnight at 37°C.
+* Check results tomorrow.