Difference between revisions of "Team:Munich/assembly2.html"
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<td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | ||
| − | <td> T4 Phage: | + | <td> T4 Phage: 4*10^5 PFU/mL |
| − | T7 Phage: 5*10^ | + | T7 Phage: 5*10^7 PFU/mL |
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<td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | <td><a href="https://2018.igem.org/Team:Munich/Results" target="_blank">Results:</a></td> | ||
| − | <td> 3S Phage: 2*10^ | + | <td> 3S Phage: 2*10^4 PFU/mL |
T4 Phage: 5*10^4 PFU/mL | T4 Phage: 5*10^4 PFU/mL | ||
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<td>Protocol:</td> | <td>Protocol:</td> | ||
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| − | <a href=" | + | <a href="https://international.neb.com/protocols/2014/10/24/rapid-pngase-f-by-sds-page-protocol-p0710" target="_blank">SDS-Page</a> |
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Latest revision as of 03:11, 18 October 2018
T7 and Phage Assembly in Commercial Cell-Extract
2018/07/31| Participants: | Brigit Tunaj |
| Protocol: | PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | T7 and T4 Phage: 2% PEG, 4% PEG, 6% PEG |
| Results: | T4 4%: 3*10^4 PFU/mL, T7 %, 1*10^8 PFU/mL |
Phage Titer from T4 and T7
2018/08/02| Participants: | Brigit Tunaj |
| Protocol: | Agar overlay plaque assay |
| Notes: | Standard procedure |
| Results: | T4 Phage: 4*10^5 PFU/mL T7 Phage: 5*10^7 PFU/mL |
T7 Phage Assembly in Commercial Cell-Extract
2018/08/08- 2018/08/09| Participants: | Sophie Kurzbach |
| Protocol: | PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | DNA from Kilian (10,37 nM) |
| Results: | No plaques |
T4 and 3S Phage Assembly in Commercial Cell-Extract
2018/08/08- 2018/08/09| Participants: | Brigit Tunaj |
| Protocol: | PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | Standard procedure |
| Results: | 3S Phage: 2*10^4 PFU/mL T4 Phage: 5*10^4 PFU/mL |
T7 Phage Burst size Analysis and Assembly in Commercial Cell-Extract
2018/08/08| Participants: | Sophie Kurzbach |
| Protocol: | Burst size analysis, PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | No notes |
| Results: | Burst size: 408 |
T7 Phage Assembly in Commercial Cell-Extract
2018/08/10- 2018/08/11| Participants: | Sophie Kurzbach |
| Protocol: | PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | DNA from Kilian (10,37 nM) |
| Results: | No plaques |
3S Phage Assembly in Commercial Cell-Extract
2018/08/07 2018/08/08| Participants: | Brigit Tunaj |
| Protocol: | PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | Standard procedure |
| Results: | No plaques |
T7 Phage Assembly in Commercial Cell-Extract
2018/08/13- 2018/08/14| Participants: | Sophie Kurzbach |
| Protocol: | PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | DNA from Kilian |
| Results: | No plaques |
| Participants: | Brigit Tunaj |
| Protocol: | PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | Standard procedure |
| Results: | 10^3 PFU/mL |
| Participants: | Sophie Kurzbach |
| Protocol: | Burst size analysis, PhageT7AssemblyTXTL, Agar overlay plaque assay |
| Notes: | Standard procedure |
| Results: | Burst size: 1250 |
Burst size with T7
2018/09/21| Participants: | Sophie Kurzbach |
| Protocol: | Burst size analysis (Heinemann, 2007), Agar overlay plaque assay | Notes: | Time for adsorption: 5 min instead of 12 min, time until lysis ≈ 1 h | Results: | Burst size synthetic T7 = 39600 PFU/mL, natural T7 = 9500 PFU/mL |
Amicon MW 100 coprecipitation of T4 phages in cell extract
2018/09/22| Participants: | Sophie Kurzbach |
| Protocol: | SDS-Page | Notes: | No notes. | Results: | DS Page: no bands (label: P (1 µL T4 + 6.5 µL ddH2O), E ((1 µL T4 + 6.5 µL ddH2O). |
Amicon MW 100 coprecipitation of T4 phages in cell extract
2018/09/24| Participants: | Sophie Kurzbach |
| Protocol: | SDS-Page | Notes: | No notes. | Results: | Denaturated phage complex in low concentration -> ROTI dye not sensitive enough to show bands specific for phages, significant band at 9 kDa for soc-protein possible. |
Burst size analysis
2018/09/25| Participants: | Sophie Kurzbach |
| Protocol: | Burst size analysis (1:10 Transfection) | Notes: | No notes. | Results: |
Natural T7 (2,2 x 10^10 PFU/mL) Synthetic T7 (13.08.2018) (2,36 x 10^11 PFU/mL) |
Plaque Assay Amicon filtered T4 phage
2018/09/26| Participants: | Sophie Kurzbach |
| Protocol: | Agar overlay plaque assay | Notes: | Used Amincon filter. | Results: | 2,05 x 10^10 PFU/mL, After: 5,24 x 10^7 PFU/mL. |
Burst size analysis T7 Phage
2018/10/01| Participants: | Sophie Kurzbach |
| Protocol: | Burst size analysis (Heinemann, 2007) | Notes: | No notes. | Results: |
Ni = Titer 1 - Titer 2 = 5.94 x 10^7 Burst size = Titer 3/Ni = 202 |
Burst size analysis
2018/10/8| Participants: | Sophie Kurzbach |
| Protocol: | Burst size analysis (Heinemann, 2007) | Notes: | With T7 Phage | Results: | burst size: 120 |