Difference between revisions of "Team:UCLA/Notebook/Recombinant Expression/21 May 2015"

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** [https://2015.igem.org/Team:UCLA/Notebook/Recombinant_Expression/20_May_2015 Yesterday's] 50mL culture of BL21(DE3) streaked from Mark Arbing and starter cultures made were grown to OD600 of 1.2 -- way above the 0.4-0.6 range recommend by Zymogen.   
 
** [https://2015.igem.org/Team:UCLA/Notebook/Recombinant_Expression/20_May_2015 Yesterday's] 50mL culture of BL21(DE3) streaked from Mark Arbing and starter cultures made were grown to OD600 of 1.2 -- way above the 0.4-0.6 range recommend by Zymogen.   
 
** Competency was conducted regardless, resuspending cell pellets in competency buffer wafter wash buffering the cells from each culture.
 
** Competency was conducted regardless, resuspending cell pellets in competency buffer wafter wash buffering the cells from each culture.
 +
** David aliquotted the resulting DH5alpha and BL21(DE3) cells into 50uL samples, and froze in -80.

Revision as of 00:24, 22 May 2015

  • Starter culture creation
    • Looked at plates streaked from [tubes Tokyo gBlock 3/4/5]
    • Each plate had a good amount of isolated colonies, well streaked.
    • Observation of the streaked colonies under gel box UV yielded several colonies with high fluorescence, enough to verify proper insert presence.
    • Colony morphology was also identical, which indicates lack of satellite colonies.
  • Zymo Mix N' Go Competency tested
    • Yesterday's 50mL culture of BL21(DE3) streaked from Mark Arbing and starter cultures made were grown to OD600 of 1.2 -- way above the 0.4-0.6 range recommend by Zymogen.
    • Competency was conducted regardless, resuspending cell pellets in competency buffer wafter wash buffering the cells from each culture.
    • David aliquotted the resulting DH5alpha and BL21(DE3) cells into 50uL samples, and froze in -80.