Difference between revisions of "User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/24"

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(Transformation of BL21 cells)
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==Transformation of BL21 cells==
 
==Transformation of BL21 cells==
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The following procedures were conducted near the proximity of the Bunsen burner in accordance with aseptic techniques. The procedure is based from the laboratory [[AU Biomaterials Design Lab:Protocols/Transformation Protocol|Transformation protocol.]]
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* Transferred 5 μL of PCR plasmid in two sterilized tubes. One tube will be filled with LB media, the other with SOC media. Both media are from Novagen.
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* Added 40 μL of thawed BL21 cells into each tube. The tubes were immediately placed on a VWR analog heatblock at 42°C for 30 sec.
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* After the 30 sec. mark, the tubes were submerged in ice. This rapid change in temperatures is a method of heat shock. This is an adequate of method of allowing the cells to incorporate the mutated plasmid into their intracellular matrix.
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* Delivered 250 μL of the LB and SOC media using an automatic delivery pipet into the labeled tubes.
 
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Revision as of 11:04, 27 October 2012

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Transformation of BL21 cells

The following procedures were conducted near the proximity of the Bunsen burner in accordance with aseptic techniques. The procedure is based from the laboratory Transformation protocol.

  • Transferred 5 μL of PCR plasmid in two sterilized tubes. One tube will be filled with LB media, the other with SOC media. Both media are from Novagen.
  • Added 40 μL of thawed BL21 cells into each tube. The tubes were immediately placed on a VWR analog heatblock at 42°C for 30 sec.
  • After the 30 sec. mark, the tubes were submerged in ice. This rapid change in temperatures is a method of heat shock. This is an adequate of method of allowing the cells to incorporate the mutated plasmid into their intracellular matrix.
  • Delivered 250 μL of the LB and SOC media using an automatic delivery pipet into the labeled tubes.