Team:University College London/LabBook/Week5
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! colspan="2" | Samples !! Volume Inoculated !! Antibiotic in Gel (ug/ml) | ! colspan="2" | Samples !! Volume Inoculated !! Antibiotic in Gel (ug/ml) | ||
|- | |- | ||
- | | rowspan="16" |BioBrick ||rowspan="2" | BBa_ K123003 || | + | | rowspan="16" |BioBrick ||rowspan="2" | BBa_ K123003 || 20ul || rowspan="14" | Ampicillin(50ug/ml) |
|- | |- | ||
| 200ul | | 200ul |
Revision as of 12:21, 1 August 2012
Contents |
Monday (9.7.12)
Aim: Transformation of Key BioBricks
Method
(LOGO) Transformation Protocol 2
Step 1 – Thawing Cells: Use W3100 cell line created in Week 2 (Expt 2.1) Step 3 – Addition of BioBrick: To each 2ml eppendorf, add 1ul of the following BioBricks. Include an extra tube as a control, with no BioBrick added.
Function | Module | ||
---|---|---|---|
BioBrick | BBa_K123003 | Oestrogen Receptor | Detection |
BBa_K123002 | Oestrogen Response Element | Detection | |
BBa_K398108 | Salt Tolerance Cluster | Salt Tolerance | |
BBa_J23100 | Constitutive Promoter | ||
BBa_J23107 | Constitutive Promoter | ||
BBa_R0040 | TetR Repressible Promoter | Buoyancy | |
BBa_J23106 | Constitutive Promoter | ||
BBa_B0034 | Ribosome Binding Site (RBS) | All | |
Control | (No BioBricks) |
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_ K123003 | 20ul | Ampicillin(50ug/ml) |
200ul | |||
BBa_ K123002 | 20ul | ||
200ul | |||
BBa_B0034 | 20ul | ||
200ul | |||
BBa_J23100 | 20ul | ||
200ul | |||
BBa_J23107 | 20ul | ||
200ul | |||
BBa_R0040 | 20ul | ||
200ul | |||
BBa_J23106 | 20ul | ||
200ul | |||
BBa_ K398108 | 20ul | Chloramphenicol (25ug/ml) | |
200ul | |||
Control | Positive (No BioBrick) | 36ul | No Antibiotic |
Negative (No BioBrick) | 36ul | 2x Ampicillin(50ug/ml)
1x Chloramphenicol (25ug/ml) |
Tuesday (10.7.12)
Aim - Check results from Transformation
Results: The table below indicates whether or not there was growth on each plate. Included is an image demonstrating the growth noted for BBa_K123003, BBa_K398108, and the Positive Control
Samples | Volume Inoculated | Growth/No Growth | |
---|---|---|---|
BioBrick | BBa_ K123003 | 20ul | Growth |
200ul | Growth | ||
BBa_ K123002 | 20ul | No Growth | |
200ul | No Growth | ||
BBa_B0034 | 20ul | No Growth | |
200ul | No Growth | ||
BBa_J23100 | 20ul | No Growth | |
200ul | No Growth | ||
BBa_J23107 | 10ul | No Growth | |
2000ul | No Growth | ||
BBa_R0040 | 20ul | No Growth | |
200ul | No Growth | ||
BBa_J23106 | 20ul | No Growth | |
200ul | No Growth | ||
BBa_ K398108 | 20ul | No Growth | |
200ul | Growth | ||
Control | Positive (No BioBrick) | 36ul | Growth |
Negative (No BioBrick) | 36ul | No Growth |
Conclusion: Only BBa_K398108 and BBa_K123003 had successfully transformed. This is a serious setback, which suggests our cell competency is not as high as we first anticipated. Method
(LOGO) Picking Colonies
Step 2 - Inoculating Colonies into a Selective Broth: The table below indicates the volume of broth and the concentration of antibiotic used for the BioBrick.
4.1
5.3
5.2