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Team:TUDelft/6 August 2009
From 2009.igem.org
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| D1 || TRI || 10<sup>0</sup> || overgrown | | D1 || TRI || 10<sup>0</sup> || overgrown | ||
|- align="center" | |- align="center" | ||
| - | | D2 || TRI || 10<sup>-2</sup>|| <font color=green><B>6770</b></font> | + | | [https://2009.igem.org/Image:TUDelft-aug5-2009-conj-test-plate-D2-noGFP.jpg D2] || TRI || 10<sup>-2</sup>|| <font color=green><B>6770</b></font> |
|- align="center" | |- align="center" | ||
| D3 || TRI || 10<sup>-4</sup>|| 170 | | D3 || TRI || 10<sup>-4</sup>|| 170 | ||
| Line 31: | Line 31: | ||
| T2 || TRI + AMP || 10<sup>-1</sup>|| ? | | T2 || TRI + AMP || 10<sup>-1</sup>|| ? | ||
|- align="center" | |- align="center" | ||
| - | | T3 || TRI + AMP || 10<sup>-2</sup>|| <font color=green><B>186</b></font> | + | | [https://2009.igem.org/Image:TUDelft-aug5-2009-conj-test-plate-T3-GFP.jpg T3] || TRI + AMP || 10<sup>-2</sup>|| <font color=green><B>186</b></font> |
|- align="center" | |- align="center" | ||
| T4 || TRI + AMP || 10<sup>-3</sup>|| 11 | | T4 || TRI + AMP || 10<sup>-3</sup>|| 11 | ||
| Line 59: | Line 59: | ||
| D6 || TRI || 10<sup>0</sup> || overgrown | | D6 || TRI || 10<sup>0</sup> || overgrown | ||
|- align="center" | |- align="center" | ||
| - | | D7 || TRI || 10<sup>-2</sup>|| <font color=green><B>8320</b></font> | + | | [https://2009.igem.org/Image:TUDelft-aug5-2009-conj-test-plate-D7-noGFP.jpg D7] || TRI || 10<sup>-2</sup>|| <font color=green><B>8320</b></font> |
|- align="center" | |- align="center" | ||
| D8 || TRI || 10<sup>-4</sup>|| 114 | | D8 || TRI || 10<sup>-4</sup>|| 114 | ||
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| T9 || TRI + AMP || 10<sup>-1</sup>|| ? | | T9 || TRI + AMP || 10<sup>-1</sup>|| ? | ||
|- align="center" | |- align="center" | ||
| - | | T10 || TRI + AMP || 10<sup>-2</sup>|| <font color=green><B>228</b></font> | + | | [https://2009.igem.org/Image:TUDelft-aug5-2009-conj-test-plate-T10-GFP.jpg T10] || TRI + AMP || 10<sup>-2</sup>|| <font color=green><B>228</b></font> |
|- align="center" | |- align="center" | ||
| T11 || TRI + AMP || 10<sup>-3</sup>|| 18 | | T11 || TRI + AMP || 10<sup>-3</sup>|| 18 | ||
| Line 91: | Line 91: | ||
| D11 || TRI || 10<sup>0</sup> || overgrown | | D11 || TRI || 10<sup>0</sup> || overgrown | ||
|- align="center" | |- align="center" | ||
| - | | D12 || TRI || 10<sup>-2</sup>|| <font color=green><B>8950</b></font> | + | | [https://2009.igem.org/Image:TUDelft-aug5-2009-conj-test-plate-D12-noGFP.jpg D12] || TRI || 10<sup>-2</sup>|| <font color=green><B>8950</b></font> |
|- align="center" | |- align="center" | ||
| D13 || TRI || 10<sup>-4</sup>|| 173 | | D13 || TRI || 10<sup>-4</sup>|| 173 | ||
| Line 103: | Line 103: | ||
| T16 || TRI + AMP || 10<sup>-1</sup>|| ? | | T16 || TRI + AMP || 10<sup>-1</sup>|| ? | ||
|- align="center" | |- align="center" | ||
| - | | T17 || TRI + AMP || 10<sup>-2</sup>|| <font color=green><B>218</b></font> | + | | [https://2009.igem.org/Image:TUDelft-aug5-2009-conj-test-plate-T17-GFP.jpg T17] || TRI + AMP || 10<sup>-2</sup>|| <font color=green><B>218</b></font> |
|- align="center" | |- align="center" | ||
| T18 || TRI + AMP || 10<sup>-3</sup>|| 31 | | T18 || TRI + AMP || 10<sup>-3</sup>|| 31 | ||
| Line 133: | Line 133: | ||
Elecroporated 10 assemblies. Experienced some arcing with assemblies 1 and 6. Assembly 6 was redone with only 2 uL of DNA added. | Elecroporated 10 assemblies. Experienced some arcing with assemblies 1 and 6. Assembly 6 was redone with only 2 uL of DNA added. | ||
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===Tim Weenink=== | ===Tim Weenink=== | ||
Revision as of 16:04, 16 August 2009
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.
