Haynes Lab:Notebook/Engineering PC-TFs/2013/02/01

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(Autocreate 2013/02/01 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
(Summary)
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==Summary==
==Summary==
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<font size=3>'''Bacterial transformation'''</font>
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* Added total volume (10.0 μL) to 50 μL chemically competent cells  (e.g., BL21) in a 2.0 mL tube.
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* Incubated on ice for 2 min., heat shock at 42°C for exactly 45 sec. (''Accidentally for 1 min''), and placed on ice.
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* Added 800 μL sterile SOC medium.
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* Grow with shaking at 37°C for 30 min.(''Was confused about how to put 2ml tubes in shaker.'') 
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* Pelleted the cells at top speed in a microcentrifuge for 3 min. at room temperature.
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* Discarded the supernatant. Resuspended the cells in 100 μL LB + antibiotic.
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* Plate cells on pre-warmed LB agar + antibiotic. (''I plated 100 μL of the cells. Protocol didn't specify.'')  Grow overnight at 37°C.
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* Check results tomorrow.
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Revision as of 22:48, 1 February 2013

Engineering PC-TFs Main project page
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Summary

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. (Accidentally for 1 min), and placed on ice.
  • Added 800 μL sterile SOC medium.
  • Grow with shaking at 37°C for 30 min.(Was confused about how to put 2ml tubes in shaker.)
  • 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. Protocol didn't specify.) Grow overnight at 37°C.
  • Check results tomorrow.



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