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Team:University College London/LabBook/Week3
From 2012.igem.org
Monday 23.7.12
Aim - Transformation of TetR BBa_C0040 BioBrick
(LOGO) Transformation Protocol 1
Step 1 – Thawing Cells: Use W3100 cell line created in Week 2 (Expt 2.1)
Step 3 – Addition of BioBrick: To a 2ml eppendorf, add 1ul of the following BioBricks. Note: we have changed the protocol for our positive control. Previously it contained no BioBrick, but it has been recommended to us that we transform our positive control such that there is one for each BioBrick – this will tell us if the BioBrick has in any way affected cell viability. This will be used from this point onwards. Include an extra tube as a negative control, with no BioBrick added
| Samples | Function | Module | |
|---|---|---|---|
| BioBrick | BBa_C0040 | Tetracycline Repressor | Buoyancy |
| Control | Positive (Contains BioBrick BBa_C0040) | ||
| Negative (No Biobrick) | |||
Step 9 - Plating samples on Agar Plates: The table below indicates the chosen inoculation volume (two for each BioBrick) and the correct gel antibiotic concentration for all samples.
| Samples | Volume Inoculated | Antibiotic in Gel (ug/ml) | |
|---|---|---|---|
| BioBrick | BBa_C0040 | 10ul | Ampicillin(50ug/ml) |
| 90ul | |||
| Control | Positive (Contains BioBrick BBa_C0040) | 36ul | No Antibiotic |
| Negative (No BioBrick) | 36ul | 2x Ampicillin(50ug/ml) | |
Tuesday 24.7.12
Aim - Results of Transformation
Result: The table below indicates that there was growth for this transformation.
| Samples | Volume Inoculated | Colony Formation | |
|---|---|---|---|
| BioBrick | BBa_C0040 | 10ul | Yes |
| 90ul | Yes | ||
| Control | Positive (Contains BioBrick BBa_C0040) | 36ul | Yes |
| Negative (No BioBrick) | 36ul | No | |
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