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| | how you used this part and how it worked out. | | how you used this part and how it worked out. |
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| − | ===Applications of BBa_K1970004=== | + | ===Applications of BBa_K1970000=== |
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| | ===User Reviews=== | | ===User Reviews=== |
| − | team:<b>NTNU 2016</b>
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| − | ===Applications of BBa_K608351===
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| − | We tested this part together with an mCherry reporter ([https://parts.igem.org/Part:BBa_E1010]) controlled by a self-inhibiting RBS. The retrons in this part are set to produce msDNA that targets the inhibitory stem loop of this RBS thereby inducing translational activity from it (an thus production of mCherry). Additionally the
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| − | E. Coli DH5α cells were transformed with the constructs above. Transformants were inoculated overnight in LB medium with kanamycin, at 37 °C and 255rpm. The 5 cultures were placed in a water bath at 37 °C. The water bath was gradually heated to 42 °C to avoid heat shocking the cells. Time courses of the fluorescence (abs: 580nm, em: 615nm) and OD of the cells were recorded (see the figure below).
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| − | [[Image:24_10_1_fused.png|900px]]
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| − | [[Image:24_10_2_fused.png|900px]]
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| − | The four combinations of RBS and terminator around an mCherry reporter coupled to a CI repressor (this part). In a) and b) the RBS was clearly not effective, with the fluorescence being close to that of cells lacking an mCherry. The heat treatment did not have any obvious influence on the fluorescence of the cells. The RBS in c) and d) had relatively high fluorescence values, which changed by an order of magnitude with the terminator chosen. However, the heat treatment did not have any obvious positive influence in these cases either. In d) the influence was negative in fact.
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| − | Therefore we concluded that this part is not a good choice for temperature transitions between 37 and 42 °C. This agrees with the experience gained on this part by other groups.
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| | ===User Reviews=== | | ===User Reviews=== |
| − | <!-- DON'T DELETE --><partinfo>BBa_K1970004 StartReviews</partinfo> | + | <!-- DON'T DELETE --><partinfo>BBa_K1970000 StartReviews</partinfo> |
| | <!-- Template for a user review | | <!-- Template for a user review |
| | {|width='80%' style='border:1px solid gray' | | {|width='80%' style='border:1px solid gray' |
| | |- | | |- |
| | |width='10%'| | | |width='10%'| |
| − | <partinfo>BBa_K1970004 AddReview number</partinfo> | + | <partinfo>BBa_K1970000 AddReview number</partinfo> |
| | <I>Username</I> | | <I>Username</I> |
| | |width='60%' valign='top'| | | |width='60%' valign='top'| |
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| | |}; | | |}; |
| | <!-- End of the user review template --> | | <!-- End of the user review template --> |
| − | <!-- DON'T DELETE --><partinfo>BBa_K1970004 EndReviews</partinfo> | + | <!-- DON'T DELETE --><partinfo>BBa_K1970000 EndReviews</partinfo> |
UNIQf03460b546956dba-partinfo-00000000-QINU
UNIQf03460b546956dba-partinfo-00000001-QINU
