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Team:BCCS-Bristol/Calendar-Notebook/13 August 2008
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(New page: ==BioBrick Transformation== The self-made electro competent ''E. coli'' DH5α cells were tested twice. The transformation efficiency with pUC19 was 4.45 x 10<sup>8</sup> cfu/µg and 5.67 x...) |
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==BioBrick Transformation== | ==BioBrick Transformation== | ||
The self-made electro competent ''E. coli'' DH5α cells were tested twice. The transformation efficiency with pUC19 was 4.45 x 10<sup>8</sup> cfu/µg and 5.67 x 10<sup>8</sup> cfu/µg. The first attempt with BioBrick DNA failed, but this might be due to a too low cell amount compared to all the other decanted tubes. In the following transformations, the incubation of the punched paper disc with the DNA was varied in time (3 h and 5.5 h). It seems that a longer incubation increases the transformation efficiency. This has to be confirmed, since the transformations were conducted by different persons. Thereby, next to the [http://partsregistry.org/Part:BBa_J63005 yeast ADH1 promoter] another BioBrick ([http://partsregistry.org/Part:BBa_J63001 enhanced version of EYFP, yeast-optimized YFP]) was tested because the VF2-VR value of [http://partsregistry.org/Part:BBa_J63005 yeast ADH1 promoter] we got didn't coincide with the value given on the iGEM page. | The self-made electro competent ''E. coli'' DH5α cells were tested twice. The transformation efficiency with pUC19 was 4.45 x 10<sup>8</sup> cfu/µg and 5.67 x 10<sup>8</sup> cfu/µg. The first attempt with BioBrick DNA failed, but this might be due to a too low cell amount compared to all the other decanted tubes. In the following transformations, the incubation of the punched paper disc with the DNA was varied in time (3 h and 5.5 h). It seems that a longer incubation increases the transformation efficiency. This has to be confirmed, since the transformations were conducted by different persons. Thereby, next to the [http://partsregistry.org/Part:BBa_J63005 yeast ADH1 promoter] another BioBrick ([http://partsregistry.org/Part:BBa_J63001 enhanced version of EYFP, yeast-optimized YFP]) was tested because the VF2-VR value of [http://partsregistry.org/Part:BBa_J63005 yeast ADH1 promoter] we got didn't coincide with the value given on the iGEM page. | ||
Revision as of 10:00, 19 August 2008
BioBrick Transformation
The self-made electro competent E. coli DH5α cells were tested twice. The transformation efficiency with pUC19 was 4.45 x 108 cfu/µg and 5.67 x 108 cfu/µg. The first attempt with BioBrick DNA failed, but this might be due to a too low cell amount compared to all the other decanted tubes. In the following transformations, the incubation of the punched paper disc with the DNA was varied in time (3 h and 5.5 h). It seems that a longer incubation increases the transformation efficiency. This has to be confirmed, since the transformations were conducted by different persons. Thereby, next to the [http://partsregistry.org/Part:BBa_J63005 yeast ADH1 promoter] another BioBrick ([http://partsregistry.org/Part:BBa_J63001 enhanced version of EYFP, yeast-optimized YFP]) was tested because the VF2-VR value of [http://partsregistry.org/Part:BBa_J63005 yeast ADH1 promoter] we got didn't coincide with the value given on the iGEM page.
