User:Mary Mendoza/Notebook/CHEM 571 Experimental Biological Chemistry I/2012/10/24

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Transformation of BL21 cells)
(Transformation of BL21 cells)
Line 13: Line 13:
* 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.
* The tubes containing the media were centrifuge for 1 h. at 37°C by Dr. Miller.
-
*  
+
* During the latter of the lab period, 2 plates were inoculated for each cell grown in LB and SOC media.
-
 
+
* The first plate was inoculated with 50 μL of the BL21 cell grown in LB; the second plate was inoculate with a greater volume of cells with 200 μL.
 +
* Dhea Patel plated the BL21 cells grown in SOC media.

Revision as of 14:12, 27 October 2012

Project name Main project page
Previous entry      Next entry

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.
  • During the latter of the lab period, 2 plates were inoculated for each cell grown in LB and SOC media.
  • The first plate was inoculated with 50 μL of the BL21 cell grown in LB; the second plate was inoculate with a greater volume of cells with 200 μL.
  • Dhea Patel plated the BL21 cells grown in SOC media.



Personal tools