Imperial College/10 August 2008
From 2008.igem.org
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* WETLAB TEAM | * WETLAB TEAM | ||
- | #Begin culturing ''B.subtilis'' | + | #Begin culturing ''B.subtilis'' and transformation: |
+ | *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, | ||
+ | |||
+ | |||
#Transform XL1-blue ''E.coli'' with Biobricks containing BBa_B0015 and BBa_c0012 | #Transform XL1-blue ''E.coli'' with Biobricks containing BBa_B0015 and BBa_c0012 | ||
#Design all primers for PCR cloning | #Design all primers for PCR cloning |
Revision as of 22:43, 7 August 2008
- WETLAB TEAM
- Begin culturing B.subtilis and transformation:
- 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,
- Transform XL1-blue E.coli with Biobricks containing BBa_B0015 and BBa_c0012
- Design all primers for PCR cloning
- PCR clone AmyE from pDR111 and XylR from the B.subtilis genome
- Prepare B.subtilis for first microscopy session - a set of dilutions to ascertain optimal amount of cells per mL
- Chris and James will attend microscope training
- Produce cell number against OD600nm calibration curve for use in characterisation
- DRYLAB TEAM
- Finish up tutorial 2 by this week
- Start working on tutorial 3
- Attend microscope training
- Generate data sets from bacteria motility movies