"
Team:TU Darmstadt/Notebook/Labjournal/K1497016
From 2014.igem.org
(Difference between revisions)
| Line 4: | Line 4: | ||
<html> | <html> | ||
| - | |||
| - | |||
| - | |||
| - | |||
| - | |||
<div id="leftNavi" class="grid_5"> | <div id="leftNavi" class="grid_5"> | ||
<nav> | <nav> | ||
| - | <ul class="menu"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt | + | <ul class="menu"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt" >Home</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Project" >Project</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Results" >Results</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/PolicyandPractices" >Policy & Practices</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Achievements" >Achievements</a></li><li class="active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook" >Notebook</a><ul class="menu2"><li class="active first"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal" >Labjournal</a><ul class="menu3"><li class="first"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497000" >K1497000</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497001" >K1497001</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497002" >K1497002</a></li><li ><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497003" >K1497003</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497004" >K1497004</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497005" >K1497005</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497006" >K1497006</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497007" >K1497007</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497008" >K1497008</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497009" >K1497009</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497010" >K1497010</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497011" >K1497011</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497012" >K1497012</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497013" >K1497013</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497014" >K1497014</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497015" >K1497015</a></li><li class="active"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497016" >K1497016</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497017" >K1497017</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497018" >K1497018</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497019" >K1497019</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497020" >K1497020</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497021" >K1497021</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497022" >K1497022</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497023" >K1497023</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497024" >K1497024/5/6</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497027" >K1497027/28/29/30</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497031" >K1497031</a></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497032" >K1497032</a></li><li class="last"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Labjournal/K1497033" >K1497033</a></li></ul></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Materials" >Materials</a></li><li class="last"><a href="https://2014.igem.org/Team:TU_Darmstadt/Notebook/Methods" >Methods</a></li></ul></li><li><a href="https://2014.igem.org/Team:TU_Darmstadt/Team" >Team</a></li><li class="last"><a href="https://2014.igem.org/Team:TU_Darmstadt/Gallery" >Gallery</a></li></ul> |
</nav> | </nav> | ||
</div> | </div> | ||
Latest revision as of 15:09, 17 October 2014
K1497016 - Constitutive Naringenin production operon
We decided to use two different kinds of promoter systems. As a constitutiv pomoter, we chose J23100 from the registry. This promoter is placed on a plasmid called J61002. We had to place our operon behind the promoter onto J61002. So we cut J23100-J61002 with SpeI + PstI and dephosphorylated the digestion. Furthermore we digested B0034-4CL-B0034-TAL-B0034-CHS-B0034-CHI-pSB1C3 (K1497007) with XbaI + PstI. We ligated both digestions with T4 ligase and transformed the products into Top10 E.coli via heat shock. We spread out on AMP-LB agar plates and incubated the plates overnight at 37°C. We analyzed obtained colonies by colony PCR and agarose gel electrophoresis.
Positive colonies were inoculated in 5 ml LB with AMP and plasmids (J23100-K1497007-J61002) were purified after incubating for 16 h at 37°C. But we still had our insert on a AMP vector. So we cut J23100-K1497007-J61002 with EcoRI + PstI and ligated this part with a previously digestion of mRFP-pSB1C3 with EcoRI +PstI. We transformed the ligation in Top10 via heat shock and spread out on CMP-LB agar plates. After incubating the plates overnight at 37°C, we analyzed white colonies by colony-PCR and agarose gel electrophoresis. Positive colonies were inoculated in 5 ml LB with CMP and plasmids were purified after incubating for 16 h at 37°C.
For further verification we send the samples to Eurofins for sequencing.
