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Team:TUDelft/5 August 2009
Lab Notebook
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5 August 2009
Sriram
Today I did the restiction along with Daniel. After lot of calculation and discussion we did it successfully. Then I prepared a gel and loaded all the restricted samples in the gel and it could be seen in below image:
Then we decided to continue the assembly tomorrow. Daniel gave me the instruction and he decided to check with how to do the GFP expression experiments.
Tim Weenink
| Well | Part | Expected Plasmid Size | Status |
| 1-8 | !D PCR products (wrong primer) | 1100 | ✖ |
| 9 | [http://www.eurogentec.com/EGT/Images/RESALES/Electrophoresis/Regular%20DNA%20Ladder/7-SmartLadder.jpg DNA Ladder] | ✔ | |
| 10 | pSB1AK3 linear | 3864 | ✔ |
| 11 | pSB4C5 linear | 3896 | ✔ |
| 12 | strong promoter | 2983 | ✔ |
| 13 | oriT-R linear | ? | ✔ |
Orr
Prepared 1 liter of LB medium, and helped Calin with the Conjugation procedure.
Calin
Made test plate for Trimpethoprim.
Ran conjugation test with R751 (TRI) as donor and DH5 α cells with GFP plasmid and AMP as recipients.
Ran three conjugations in parallel, with a conjugation time of 1 hour.
Starting OD for recipients 0.502, for donors 0.542.
Made a total of 40 plates:
| Set 1 | |||
| Plate ID | Antibiotics | Dilution | |
| D1 | TRI | 100 | |
| D2 | TRI | 10-2 | |
| D3 | TRI | 10-4 | |
| D4 | TRI | 10-5 | |
| D5 | TRI | 10-6 | |
| T1 | TRI + AMP | 100 | |
| T2 | TRI + AMP | 10-1 | |
| T3 | TRI + AMP | 10-2 | |
| T4 | TRI + AMP | 10-3 | |
| T5 | TRI + AMP | 10-4 | |
| T6 | TRI + AMP | 10-5 | |
| T7 | TRI + AMP | 10-6 | |
| R1 | AMP | 100 | |
| R2 | AMP | 10-4 | |
| R3 | AMP | 10-5 | |
| R4 | AMP | 10-6 |
| Set 2 | |||
| Plate ID | Antibiotics | Dilution | |
| D6 | TRI | 100 | |
| D7 | TRI | 10-2 | |
| D8 | TRI | 10-4 | |
| D9 | TRI | 10-5 | |
| D10 | TRI | 10-6 | |
| T8 | TRI + AMP | 100 | |
| T9 | TRI + AMP | 10-1 | |
| T10 | TRI + AMP | 10-2 | |
| T11 | TRI + AMP | 10-3 | |
| T12 | TRI + AMP | 10-4 | |
| T13 | TRI + AMP | 10-5 | |
| T14 | TRI + AMP | 10-6 |
| Set 3 | |||
| Plate ID | Antibiotics | Dilution | |
| D11 | TRI | 100 | |
| D12 | TRI | 10-2 | |
| D13 | TRI | 10-4 | |
| D14 | TRI | 10-5 | |
| D15 | TRI | 10-6 | |
| T15 | TRI + AMP | 100 | |
| T16 | TRI + AMP | 10-1 | |
| T17 | TRI + AMP | 10-2 | |
| T18 | TRI + AMP | 10-3 | |
| T19 | TRI + AMP | 10-4 | |
| T20 | TRI + AMP | 10-5 | |
| T21 | TRI + AMP | 10-6 |
