User:Christine Doan/Notebook/Identifying Kinase Substrates of PfCK2/2014/03/14

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new CK2

  • Induction will be done at 2pm
  • Take 2 500ul total induced samples, spin, wash, spin, resuspend
  • Spin 10min 6K 4C in large Oakridge bottle, discard LB
  • Wash in 25ml MK buffer, transfer to small Oakridge tube
  • Spin 10min 6K 4C, discard MK buffer
  • Resuspend in 30ml MK buffer + 300ul 100x protease inhibitor
  • Store 5ml in a conical in -80C
  • Store 25ml in a conical in -80C

Substrates

  • Bad news: did PCR cleanups wrong, yielded very low concentrations
  • Good news: calculations prove that there is enough PCR product to do T4 treatment for cloning
  • Bad news: the pET-30 kit has not come in yet
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