10 August 2008

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 XL1-blue 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 OD_600nm calibration curve for use in characterisation

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, BBa_C0012 and pSB1A3

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
• Chris, James, Prudence and Clinton will attend microscope training

Drylab Team

End of week five

Finish tutorial 2:

• Maximum Likelihood Estimation
• Moments
• The Bayesian Approach

Next week

• Start working on tutorial 3 (hypothesis testing)
• Generate data sets from bacteria motility movies

Thursday

• Attend microscope training (Clinton and Prudence)