User:Tamra L. Fisher/Notebook/Research for Dr. Hartings/2012/03/11

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Miniprep of Transformed NovaBlue E.coli

  • In a sterile test tube with metal cap 10mL of LB with 100μg/mL of ampicillin and one colony from the transformation plate (Friday 3/9/12, PCR product from 3/7/12) were grown overnight (~17 hours). Three of these cultures were prepared.
  • Also, the transformation of wt swMb pT7-7100mL into NovaBlue (2/27/12) was grown overnight in 100mL of LB with 100μg/mL of ampicillin. The original plan was to do a midiprep of this plasmid, but due to lack of available equipment at the time a miniprep was done instead, so only 10mL of this culture was used.
  • The four cultures were miniprepped using this procedure: Wizard® Plus SV Minipreps DNA Purification System
    • The Centrifuge Protocol was used
  • The three plasmids from PCR (3/7/12) will be sent off for sequencing at the Johns Hopkins Synthesis & Sequencing Facility



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