Registry/Measurement kit/Notebook/2007-8-3

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==E0240==
*Trying mutagenic PCR of E0240.
*Trying mutagenic PCR of E0240.
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*Used 17 cycles. Don't know what # is optimal. Stratagene recommends 12, but people seem to use ~18.
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**Too many cycles -> more junk DNA.
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*Digesting with [[DpnI]]. Felix suggested digesting for >2h.
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==Transformations==
*Later will try re-transforming into another set of competent Top10 cells.
*Later will try re-transforming into another set of competent Top10 cells.
*Should make more MG1655.
*Should make more MG1655.
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*Re-tried yesterday's transformations into Top10s.

Revision as of 16:41, 3 August 2007

E0240

  • Trying mutagenic PCR of E0240.
  • Used 17 cycles. Don't know what # is optimal. Stratagene recommends 12, but people seem to use ~18.
    • Too many cycles -> more junk DNA.
  • Digesting with DpnI. Felix suggested digesting for >2h.

Transformations

  • Later will try re-transforming into another set of competent Top10 cells.
  • Should make more MG1655.
  • Re-tried yesterday's transformations into Top10s.
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