User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/24: Difference between revisions
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==Transformation of BL21 cells== | ==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 [[AU Biomaterials Design Lab:Protocols/Transformation Protocol|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. | |||
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Revision as of 12:04, 27 October 2012
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Transformation of BL21 cellsThe 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.
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