Team:Stockholm/11 September 2010
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+ | ===Preparation of Top10 chemically competent cells=== | ||
+ | Picked 8 single colonies from the 10/9 LB plate with pipette tips. Colonies were restreaked onto an LB agar plate with Amp 100 to verify that they were not contaminated, and the pipette tips were then used to inoculate cultures of 5 ml LB. Cultures grown for 40 h in 30 °C w/o rotary shaking. | ||
+ | |||
+ | ===Extraction of RBS BioBrick (BBa_B0034)=== | ||
+ | |||
+ | ====Colony PCR==== | ||
+ | Picked 4 colonies (RBS 1-4) from the 10/9 ON plate and ran a colony PCR. | ||
+ | *Positive control (PC): pSB1C3.RFP | ||
+ | *Standard colony PCR protocol | ||
+ | **Elongation: 0:45 | ||
+ | |||
+ | ====Gel verification==== | ||
+ | 1.5 % agarose, 90 V | ||
+ | |||
+ | '''Expected bands''' | ||
+ | *RBS 34: 250 bp | ||
+ | *PC: 1385 bp |
Revision as of 17:35, 13 September 2010
Contents |
Andreas
Preparation of Top10 chemically competent cells
Picked 8 single colonies from the 10/9 LB plate with pipette tips. Colonies were restreaked onto an LB agar plate with Amp 100 to verify that they were not contaminated, and the pipette tips were then used to inoculate cultures of 5 ml LB. Cultures grown for 40 h in 30 °C w/o rotary shaking.
Extraction of RBS BioBrick (BBa_B0034)
Colony PCR
Picked 4 colonies (RBS 1-4) from the 10/9 ON plate and ran a colony PCR.
- Positive control (PC): pSB1C3.RFP
- Standard colony PCR protocol
- Elongation: 0:45
Gel verification
1.5 % agarose, 90 V
Expected bands
- RBS 34: 250 bp
- PC: 1385 bp