Team:TUDelft/5 August 2009
Lab Notebook
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5 August 2009
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 |