Team:University of Ottawa/15 July 2008

From 2008.igem.org

Contents

Today on the Lab

Tammy

Transformation of XL10 Competent E. coli cells with pDR197:AtCKX2
  • 1 uL Pure pDR197:AtCKX2
  • 1 ul 1:10 dilution pDR197:AtCKX2
  • 1 ul 1:100 dilution pDR197:AtCKX2
  • 100 ul XL10 E.Coli per reaction
  • 900 ul LB Broth per reaction
  • 3 LB Ampicillin (50 ug/mL) Plates
  • Began Incubation of transformed cells at 1:50 pm.

    • Matt

    Inoculation of 97 - BY4742 cells
  • I inoculated the 97 BY4742 cells once again for glycerol stock tomorrow
    • PCR confirmation
  • Second PCR confirmation with F58, F59 was performed and gave the correct band but also gave another weird band that I don't know what it is.
    • Digestion of PTP2
  • A digestion was performed on the PTP2 amplification product to create sticky ends using BamHI and xhoI and I left it in the PCR machine overnight.

    • Chris

    Digestion of AtCRE
  • AtCRE was digested using EagI at 37 C for about one hour
  • the result was run on a 1% gel, which showed two bands, both of which were expected
    • Gel Extraction of AtCRE
  • the AtCRE was extracted from the gel using the gel extraction kit and protocol
  • unfortunately, the final AtCRE sample was vacuumed up with the other waste by accident
    • Digestion of AtCRE
  • AtCRE was digested again using the same protocols as above, except that it was run on the "digest and denature" program on the thermal cycler. It was left overnight.
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