Team:HIT-Harbin/Experiments
From 2013.igem.org
Line 53: | Line 53: | ||
</div> | </div> | ||
<div class="multi-columns"> | <div class="multi-columns"> | ||
- | <h4 class="hashed"><span> | + | <h4 class="hashed"><span>1.Test of device</span></h4> |
<div class="half"> | <div class="half"> | ||
- | <p> | + | <p></p>1)Preparation of IPTG solution: add 240mg IPTG powder into 10mL dd H2O. We filtrated the solution to sterilize it and broke it into EP tubes. The concentration is 24mg/mL (100mM/mL). then we stored them in -20℃. |
+ | <p></p>2)To test the four different combination according to the strength of promoters, we made different concentrations of IPTG for the bacteria. | ||
+ | <div align="center"> | ||
+ | <p> Table 1 IPTG formula</p> | ||
+ | <img src="https://static.igem.org/mediawiki/2013/a/a9/Experiment4.png"/> | ||
+ | </div> | ||
+ | <p></p>We grew the IPTG-added culture in 37℃ at 120rpm, overnight. Unfortunately, we haven’t observed the expected color of RFP (Red Fluorescence Protein). | ||
+ | <div align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2013/c/c2/Experiment5.png"/> | ||
+ | <p>Fig 3. No red after night</p> | ||
+ | </div> | ||
+ | <p></p>3)After that, we tested the reporting sub-circuit of our device. We connect the constitutive promoter PLac with RBS+RFP+T, transformed the bacteria and grew it overnight. The red in the results proved that the reporting sub-circuit is working. | ||
+ | <div align="center"> | ||
+ | <img src="https://static.igem.org/mediawiki/2013/1/10/Experiment6.png"/> | ||
+ | <p></p>Fig 4. Constitutive promoter expressing RFP | ||
+ | <p></p>(Left is 12h, right is 24h) | ||
+ | </div> | ||
+ | <p></p>4)We sent our whole device to companies for sequencing, but it failed. | ||
+ | |||
</div> | </div> | ||
<div class="half last"> | <div class="half last"> |
Revision as of 05:24, 27 September 2013
Project/experiments
Experiment Schedule
![](https://static.igem.org/mediawiki/2013/4/45/Experiment2.png)
Fig 1. PCR resuLts of our own parts
(1: hrpL; 2: hrpS; 3: hrpR; 4: Ptet +strong RBS+hrpS+T; 5:Ptet+intermediate RBS+hrpS+T; 6: Ptet+weak RBS+hrpS+T; 7: PIPTG +strong RBS+hrpR+T; 8 is not needed; 9: PIPTG +weak RBS+hrpR+T; 10: PhrpL+strong RBS+tetR+T; 11: PhrpL+intermediate RBS+tetR+T; 12: PhrpL+weak RBS+tetR+T; 13: PhrpL+strong RBS+RFP+T; 14: PhrpL+weak RBS+RFP+T; 15: PhrpL+weak RBS+RFP+T)
![](https://static.igem.org/mediawiki/2013/c/ca/Experiment3.png)
Fig 1.EcoR1 and Pst1 double restriction enzyme cleavage for hrpL AND gate(BBa_K1014014) device and B-POM1(BBa_K1014999)
1:plasmid carrying BBa_K1014014; 2:double restriction enzyme cleavage for 1; 3:plasmid carrying BBa_K1014999; 4:double restriction enzyme cleavage for 3
1.Test of device
Table 1 IPTG formula
![](https://static.igem.org/mediawiki/2013/a/a9/Experiment4.png)
![](https://static.igem.org/mediawiki/2013/c/c2/Experiment5.png)
Fig 3. No red after night
![](https://static.igem.org/mediawiki/2013/1/10/Experiment6.png)
Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum.
The circuit
Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum.
Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum.
The circuit
Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum.
Lorem ipsum dolor sit amet, consectetur adipisicing elit, sed do eiusmod tempor incididunt ut labore et dolore magna aliqua. Ut enim ad minim veniam, quis nostrud exercitation ullamco laboris nisi ut aliquip ex ea commodo consequat. Duis aute irure dolor in reprehenderit in voluptate velit esse cillum.