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

From OpenWetWare
Jump to: navigation, search
(Transformation of BL21 cells)
(Transformation of BL21 cells)
Line 12: Line 12:
 
* 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.
 
* 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.  
 
* Delivered 250 μL of the LB and SOC media using an automatic delivery pipet into the labeled tubes.  
 +
* The tubes containing the media were centrifuge for 1 h. at 37°C by Dr. Miller.
 
*  
 
*  
 +
  
  

Revision as of 11:07, 27 October 2012

Owwnotebook icon.png Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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.
  • The tubes containing the media were centrifuge for 1 h. at 37°C by Dr. Miller.