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Team:Warsaw/Calendar-Main/7 July 2009
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Revision as of 20:07, 8 July 2009
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Yet another insertion of the mgtc gene into the pKSII+ plasmid
Kamil
Tasks:
- Plasmid assembly
Methods:
The ligation mix was prepared as follows: 1ul plasmid, 3ul gene, 1ul ligation buffer B (Fermentas), 1ul T4 DNA ligase (Fermentas), 1ul 30% PEG, 1ul 10mM ATP, the solution was topped up with H2O to the final volume of 10 ul. The ligation was carried out in 37°C for 1h and then inactivated for 15min. at 65°C.
A 200ul batch of chemocompetent bacteria was transformed with 3ul of ligation mix and incubated on petri dishes containing LB medium supplemented with ampicilin, X-gal and IPTG.
UPDATE: No luck this time, back to the drawing board...
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