Registry/Measurement kit/Notebook/2007-7-31

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Contents

E0240

  • Noticed that during the scarring PCR, a point substitution was made.
  • Decided to try a site-directed mutagenesis on the biobricked product.
  • Ordered two pairs of primers:
    • First pair - mutation in the ends of the primers:
      • CGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTGTAACAGC, E0240_mut_E_F (78.8)
      • CTCTCTTTTCGTTGGGATCTTTCGAAAGGGCAGATTGTGTGGACAGG, E0240_mut_E_R (79.7)
    • Second pair - mutation in the middle of the primers:
      • CCTTTCGAAAGATCCCAACGAAAAGAGAGACCACATGGTCCTTCTTGAGTTTG, E0240_mut_M_F (80)
      • ACCATGTGGTCTCTCTTTTCGTTGGGATCTTTCGAAAGGGCAGATTG, E0240_mut_M_R (79.3)

I2057

  • Re-transformed both versions with ccdB into DB3.1

I2055 and I2056

  • Letting biobrick PCR run overnight.

T9002

  • Re-tried, transformed ligation of F2620.

Misc

  • Made chemically competent DB3.1 cells.
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