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Team:TUDelft/6 August 2009
From 2009.igem.org
Lab Notebook
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6 August 2009
Calin
Good results on the conjugation test. Many colonies. All plates were imaged on the safe imager with the 5 Megapixel camera. The plates with GFP (T and R) were excited by blue light and placed under a filter (50 ms exposure). The plates without GFP (D) were imaged with the upper white light and an exposure of 15ms. Images were color inverted before the image analysis. Colony counting was done with [http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1770926 Clono-Counter].
| Set 1 | |||
| Plate ID | Antibiotics | Dilution | # Colonies |
| D1 | TRI | 100 | overgrown |
| D2 | TRI | 10-2 | 6770 |
| D3 | TRI | 10-4 | 170 |
| D4 | TRI | 10-5 | 40 |
| D5 | TRI | 10-6 | 10 |
| T1 | TRI + AMP | 100 | ? |
| T2 | TRI + AMP | 10-1 | ? |
| T3 | TRI + AMP | 10-2 | 186 |
| T4 | TRI + AMP | 10-3 | 11 |
| T5 | TRI + AMP | 10-4 | 0 |
| T6 | TRI + AMP | 10-5 | 0 |
| T7 | TRI + AMP | 10-6 | 0 |
| R1 | AMP | 100 | overgrown |
| R2 | AMP | 10-4 | 32 |
| R3 | AMP | 10-5 | 6 |
| R4 | AMP | 10-6 | 2 |
| Set 2 | |||
| Plate ID | Antibiotics | Dilution | |
| D6 | TRI | 100 | overgrown |
| D7 | TRI | 10-2 | 8320 |
| D8 | TRI | 10-4 | 114 |
| D9 | TRI | 10-5 | 18 |
| D10 | TRI | 10-6 | 8 |
| T8 | TRI + AMP | 100 | ? |
| T9 | TRI + AMP | 10-1 | ? |
| T10 | TRI + AMP | 10-2 | 228 |
| T11 | TRI + AMP | 10-3 | 18 |
| T12 | TRI + AMP | 10-4 | 2 |
| T13 | TRI + AMP | 10-5 | 0 |
| T14 | TRI + AMP | 10-6 | 0 |
| Set 3 | |||
| Plate ID | Antibiotics | Dilution | |
| D11 | TRI | 100 | overgrown |
| D12 | TRI | 10-2 | 8950 |
| D13 | TRI | 10-4 | 173 |
| D14 | TRI | 10-5 | 22 |
| D15 | TRI | 10-6 | 1 |
| T15 | TRI + AMP | 100 | ? |
| T16 | TRI + AMP | 10-1 | ? |
| T17 | TRI + AMP | 10-2 | 218 |
| T18 | TRI + AMP | 10-3 | 31 |
| T19 | TRI + AMP | 10-4 | 2 |
| T20 | TRI + AMP | 10-5 | 0 |
| T21 | TRI + AMP | 10-6 | 0 |
Calculated conjugation efficiencies:
| Set | Conjugation Efficiency |
| 1 | 0.028 |
| 2 | 0.028 |
| 3 | 0.025 |
Trimethoprim control plate had no growth.
5mL tube of assembly CA made.
Elecroporated 10 assemblies. Experienced some arcing with assemblies 1 and 6. Assembly 6 was redone with only 2 uL of DNA added.
Tim Weenink
Elecroporated 10 assemblies.
| Biobrick | origin of parts |
Orr
Made 2 one liter batches of LB medium, and prepared some chemically competent TSS cells.
