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?

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

  1. Prepare B.subtilis for first microscopy session on thursday - a set of dilutions to ascertain optimal amount of cells per mL
  2. 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
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