Difference between revisions of "Team:Jilin China/Result/Version 2"
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<p>Heat-repressible RNA-based thermosensors were based on the RNase E. We measured these thermosensors' activity by using measurement device like before. But this time, we designed a new negative control. In the following, it will be called negative control-2. Negative control-2 has a cleavage site of RNase E. It will always be digest by enzyme. Due to the effiency of RNase E, we decided to use negative control-2 instead of traditional negative control.</p> | <p>Heat-repressible RNA-based thermosensors were based on the RNase E. We measured these thermosensors' activity by using measurement device like before. But this time, we designed a new negative control. In the following, it will be called negative control-2. Negative control-2 has a cleavage site of RNase E. It will always be digest by enzyme. Due to the effiency of RNase E, we decided to use negative control-2 instead of traditional negative control.</p> | ||
<p>We designed 100 heat-repressible RNA-based thermosensors, their sequences are different. After measurement, we removed some devices which show less sensing in temperature or have undesirable results, then we finally selected 23 out of 100 heat-repressible RNA-based thermosensor in toolkit.</p> | <p>We designed 100 heat-repressible RNA-based thermosensors, their sequences are different. After measurement, we removed some devices which show less sensing in temperature or have undesirable results, then we finally selected 23 out of 100 heat-repressible RNA-based thermosensor in toolkit.</p> | ||
| − | <h4 class="tables"> | + | <h4 class="tables">·Activities of thermosensors decrease at elevated temperature</h4> |
<p>We measured the activities of these thermosensors at 3 temperatures: 29, 37 and 42℃. <b>Figure 1</b> shows the measurement results of the 23 different thermosensors. Compared with the positive control, all the heat-repressible RNA-based thermosensors' normalized fluorescence decrease at elevated temperature. They also have different intensity and sensitivity. We have added the charaterize result to the parts registry, users can choose their appropriate thermosensors.</p> | <p>We measured the activities of these thermosensors at 3 temperatures: 29, 37 and 42℃. <b>Figure 1</b> shows the measurement results of the 23 different thermosensors. Compared with the positive control, all the heat-repressible RNA-based thermosensors' normalized fluorescence decrease at elevated temperature. They also have different intensity and sensitivity. We have added the charaterize result to the parts registry, users can choose their appropriate thermosensors.</p> | ||
<img src="https://static.igem.org/mediawiki/2018/c/cc/T--Jilin_China--result--REbar.png" width="95%" length="95%"></img> | <img src="https://static.igem.org/mediawiki/2018/c/cc/T--Jilin_China--result--REbar.png" width="95%" length="95%"></img> | ||
Revision as of 08:38, 15 October 2018
VERSION 2.0
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Results
After the integrated human practice work, we update SynRT Toolkit to version 2.0. In version 2.0, we added 23 heat-repressible RNA-based thermosensors to Toolkit.
Heat-repressible RNA-based thermosensors were based on the RNase E. We measured these thermosensors' activity by using measurement device like before. But this time, we designed a new negative control. In the following, it will be called negative control-2. Negative control-2 has a cleavage site of RNase E. It will always be digest by enzyme. Due to the effiency of RNase E, we decided to use negative control-2 instead of traditional negative control.
We designed 100 heat-repressible RNA-based thermosensors, their sequences are different. After measurement, we removed some devices which show less sensing in temperature or have undesirable results, then we finally selected 23 out of 100 heat-repressible RNA-based thermosensor in toolkit.
·Activities of thermosensors decrease at elevated temperature
We measured the activities of these thermosensors at 3 temperatures: 29, 37 and 42℃. Figure 1 shows the measurement results of the 23 different thermosensors. Compared with the positive control, all the heat-repressible RNA-based thermosensors' normalized fluorescence decrease at elevated temperature. They also have different intensity and sensitivity. We have added the charaterize result to the parts registry, users can choose their appropriate thermosensors.
We also computed the fold-change from 29 to 37℃ and 37 to 42℃. As the figure shows. These fold-changes were lower than positive control and the fluorescence per Abs600 were higher than negative control-2.
Discussion: as these experiment results show, the heat-repressible RNA-based thermosensors work really well. The activities decrease with temperature increases. Additionally, the difference in fluorescence intensity and the rate of decrease points to the diversity in thermosensor response. We think the sequence change in stem length, loop size, and mismatched or bulges in the stem cause the differences in thermosensor response.
The individual heat-repressible RNA-based thermosensor's characterized results are showed as followed: