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Team:Warsaw/Calendar-Main/4 August 2009
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*The plasmid digest mix contained: 4μl purified plasmid, 2μl Tango buffer (Fermentas), 0.5μl SpeI enzyme, 0.5μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl. | *The plasmid digest mix contained: 4μl purified plasmid, 2μl Tango buffer (Fermentas), 0.5μl SpeI enzyme, 0.5μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl. | ||
*The mgtc promoter digest mix contained: 12μl purified promoter, 2μl Tango buffer (Fermentas), 0.5μl SpeI enzyme, 0.5μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl. | *The mgtc promoter digest mix contained: 12μl purified promoter, 2μl Tango buffer (Fermentas), 0.5μl SpeI enzyme, 0.5μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl. | ||
| - | *The digest was carried out in 37°C for 3h, but CIAP enzyme was added 1h before end to mgtc promoter digest mix 1h before the end of incubation. Then enzymes were inactivated in 80°C for 20min. | + | *The digest was carried out in 37°C for 3h, but 1μl CIAP enzyme was added 1h before end to mgtc promoter digest mix 1h before the end of incubation. Then enzymes were inactivated in 80°C for 20min. |
Revision as of 09:39, 4 August 2009
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Cloning of the mgtc promoter into the pSB1A3 plasmid
Monika
Tasks
- Plasmid assembly
Methods
- The plasmid digest mix contained: 4μl purified plasmid, 2μl Tango buffer (Fermentas), 0.5μl SpeI enzyme, 0.5μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl.
- The mgtc promoter digest mix contained: 12μl purified promoter, 2μl Tango buffer (Fermentas), 0.5μl SpeI enzyme, 0.5μl XbaI enzyme, the solution was topped up with H2O to the final volume of 20μl.
- The digest was carried out in 37°C for 3h, but 1μl CIAP enzyme was added 1h before end to mgtc promoter digest mix 1h before the end of incubation. Then enzymes were inactivated in 80°C for 20min.
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