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Team:DTU-Denmark/Notebook/16 July 2013
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| - | * | + | * Plasmid isolation as described in the sigma MiniPrep kit. |
* PCR in order to amplify AMO gene using USER primers and 2uL of liquid culture of ''Nitrosomonas europea''. Primers 17a and 17b were used and master mix for 5 samples was prepared. PCR program was the same as on [[Team:DTU-Denmark/Notebook/12_July_2013| 12-07-2013]] for AMO. | * PCR in order to amplify AMO gene using USER primers and 2uL of liquid culture of ''Nitrosomonas europea''. Primers 17a and 17b were used and master mix for 5 samples was prepared. PCR program was the same as on [[Team:DTU-Denmark/Notebook/12_July_2013| 12-07-2013]] for AMO. | ||
Revision as of 13:37, 17 July 2013
Contents |
208
Main purpose
- Miniprep of biobrick transformants pSB1T3, pSB3T5, pSB1AK3, pSB1A3 from 10-07-2013
- gel analysis of the colony PCRs
- PCR on AMO with USER-primers with lower temperature to see if we can get any product at all
- Order better primers for colony PCR
- PCR to extract the AMO with USER primers using Nitrosomonas cells as a template
Who was in the lab
Kristian, Gosia, Henrike, Julia
Procedure
- Plasmid isolation as described in the sigma MiniPrep kit.
- PCR in order to amplify AMO gene using USER primers and 2uL of liquid culture of Nitrosomonas europea. Primers 17a and 17b were used and master mix for 5 samples was prepared. PCR program was the same as on 12-07-2013 for AMO.
Conclusion
Nothing was on the colony-PCR gel and we speculate that this is because the USER-primers used to this are not very good to this purpose. We therefore ordered new primers for colony PCR which contain no uracil. The new primers can be used with PHUSION polymerase or any other cheaper alternative.
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