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Team:TUDelft/31 July 2009
From 2009.igem.org
Lab Notebook
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31 July 2009
Tim Weenink
Did some minipreps today. Then nanodropped them:
| Biobrick name | Internal reference | Concentration ng/µl | 260/280 | 260/230 |
| BBa_K142205 | *S2 | 86.4 | 2.01 | 2.03 |
| BBa_K142205 | *S3 | 107.7 | 2.03 | 2.11 |
| BBa_K142205 | *S4 | 69.0 | 1.97 | 1.96 |
| BBa_K142205 | *S5 | 87.4 | 1.98 | 1.99 |
| BBa_K142202 | *T1 | 128.5 | 1.99 | 2.09 |
| BBa_K142202 | *T2 | 129.4 | 2.02 | 2.16 |
| BBa_K142202 | *T3 | 126.6 | 1.97 | 2.06 |
| BBa_K142202 | *T4 | 106.7 | 1.95 | 2.05 |
| N/A | !A | 171.2 | 1.98 | 2.11 |
Calin
Cultures in all pSB4C5 tubes. Did miniprep on all tubes and also made a plate.
| Part | Concentration ng/µl | 260/280 | 260/230 |
| pSB4C5 I | 123.0 | 1.91 | 2.06 |
| pSB4C5 II | 125.8 | 1.92 | 2.29 |
| pSB4C5 III | 130.7 | 1.95 | 2.07 |
Todays gel confirmed that oriT-R digestion failed and pSB4C5 is way too large.
| Well | Part | Expected Plasmid Size | Status |
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| 22 | [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] | ✔ | |
| 23 | oriT E + P digest | 2079 | ✖ |
| 24 | pSB4C5 I | 3896 | ✖ |
| 25 | pSB4C5 II | 3896 | ✖ |
| 26 | pSB4C5 III | 3896 | ✖ |
R751 cells arrived. Made two plates without antibiotics.
Digestions were started:
| position | parts | volume in µl |
| Upstream | BBa_I13522 pTet-GFP | 27.0 |
| H2O | 15.5 | |
| Upstream | BBa_E0840 rbs-GFP-term | 5.0 |
| H2O | 37.5 | |
| Backbone | pSB1C3 | 5.0 |
| H2O | 37.5 |
