Imperial College/10 August 2008

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Contents

WETLAB TEAM

B.subtilis

Team members - James and Krupa + more if work load is too much.

Monday

  • Prepare an overnight culture of DL1-blue E.coli with the pDR110 integration vector,
  • Prepare an 2x overnight culture of B.subtilis,
  • Prepare the reagents required for transformation,

Tuesday

  • Miniprep of the DL1-blye overnight culture,
  • Make competent cells for both of the transformation protocols

Wednesday

  • Perform transformation of the competent cells using both protocols,

Thursday

  • Check successfulness of the transformation. If successful then perform transformation of linear DNA using previously made competent cells.

Friday

  • Check successfulness of the linear DNA transformation,

Produce cell number against OD600nm calibration curve for use in characterisation Do we want to do this next week or the following week? A possible activity for Friday if Wednesday's transformation is found to have failed?

Cloning

Chris and Tom for this week

Monday

  • Design all primers for PCR cloning

Tuesday

  • Transform XL1-blue E.coli with Biobricks containing BBa_B0015 and BBa_C0012

Wednesday to Friday

  • PCR clone AmyE from pDR111 and XylR from the B.subtilis genome when primers arrive
  • Prepare ALL remaining Protocols
  • Depending on arrival of primers, connect XylR and BBa_C0012 to BBa_B0015 (separately) to begin construction of inducible promoters

Microscope

  • Prepare B.subtilis for first microscopy session on thursday - a set of dilutions to ascertain optimal amount of cells per mL
  1. Chris, James, Prudence and Clinton will attend microscope training

DRYLAB TEAM

  1. Finish up tutorial 2 by this week
  2. Start working on tutorial 3
  3. Attend microscope training
  4. Generate data sets from bacteria motility movies