Team:Uppsala/InterlabStudy

From 2014.igem.org

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document.getElementById("tab1").innerHTML = '<p><h2>The study</h2><p>“All iGEM teams are invited and encouraged to participate in the first international interlab measurement study in synthetic biology. …...The goal of the interlab study is to obtain fluorescence data for three specific genetic devices expressing GFP from iGEM teams around the world.”<br<br><a href="https://2014.igem.org/Tracks/Measurement/Interlab_study">More info:https://2014.igem.org/Tracks/Measurement/Interlab_study</a></p><h3>Devices measured</h3><p>The following constructs were measured for the interlab study. Each construct was given a construct ID to make references more easy.<table id="partsT" style="width:100%"><tr><th>Construct ID</th><th>Construct</th></tr><tr><td>142</td><td>J23101-B0032-GFP(provided)</td></tr><tr> <td>177</td><td>J23101-B0032-GFP(constructed)</td></tr><tr><td>178</td><td>J23115-B0032-GFP</td></tr><tr><td>181</td><td>J23106-B0032-GFP</td></tr><tr><td>224</td><td>J23116-B0032-GFP</td></tr><tr><td>226</td><td>J23117-B0032-GFP</td></tr><tr><td>180</td> <td>J23105-B0032-GFP</td></tr><tr><td>187</td><td>J23101-B0034-BFP</td></tr><tr><td>188</td><td>J23115-B0034-BFP</td></tr></p>';
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document.getElementById("tab1").innerHTML = '<p><h2>The study</h2><p>“All iGEM teams are invited and encouraged to participate in the first international interlab measurement study in synthetic biology. …...The goal of the interlab study is to obtain fluorescence data for three specific genetic devices expressing GFP from iGEM teams around the world.”<br<br><a href="https://2014.igem.org/Tracks/Measurement/Interlab_study">More info:https://2014.igem.org/Tracks/Measurement/Interlab_study</a></p><h3>Devices measured</h3><p>The following constructs were measured for the interlab study. Each construct was given a construct ID to make references more easy.<table id="partsT" style="width:100%"><tr><th>Construct ID</th><th>Construct</th></tr><tr><td>142</td><td>J23101-B0032-GFP(provided)</td></tr><tr> <td>177</td><td>J23101-B0032-GFP(constructed)</td></tr><tr><td>178</td><td>J23115-B0032-GFP</td></tr><tr><td>181</td><td>J23106-B0032-GFP</td></tr><tr><td>224</td><td>J23116-B0032-GFP</td></tr><tr><td>226</td><td>J23117-B0032-GFP</td></tr><tr><td>180</td> <td>J23105-B0032-GFP</td></tr><tr><td>187</td><td>J23101-B0034-BFP</td></tr><tr><td>188</td><td>J23115-B0034-BFP</td></tr></table><h3>Experimental detail</h3><br><br><b>Construction of devices</b><br><br><u>DNA samples from kit:</u><br><br><p>A DNA sample of BBa_E0240 ,BBa_J23101, BBa_J23105, BBa_J23106, BBa_K823005, BBa_J23116, BBa_J23117  were taken up from the 2014 Distribution according to “Instructions on iGEMs webpage”. They were transformed into CaCl2 DH5-alpha competent cells and plated on LB-agar plates with chloramphemnicol left to incubate at 37 degrees celsius for 18h. Two clones were re-streaked on LB-agar plates with chloramphemnicol and left to incubate for 18 hours. pSB1K3 with red-insert from 2013 years distribution was re-streaked from the -80 degree cell stock.  Single colonies were PCR-amplified with VF2 and VR primers according to protocol for “Red Taq” by “ThermoFisher Scientific”. Bands were confirmed using gel electrophoresis. An overnight culture for one verified clone for each construct was prepared. Plasmid preparations were done using “Gene elute plasmid Miniprepp” following attached protocol. Concentrations of DNA were measured using “nanodrop” at 260nm.</p><h3>Assembly of devices</h3><p>All devices were constructed with B0032-GFP(BBa_E0240), and pSB1K3(from 2013 years distribution) combined via 3A assembly protocol, with HF restriction enzymes, cutsmart buffer and ligase provided from NEB, with each of the respective promoters to form the devices shown below. The two constructs with blue fluorescent protein (BFP) were constructed with B0034-BFP(BBa_K592024) instead of BBa_E0240. The assembled parts were transformed, overnighted and screened as explained above. The overnight cultures were stored in -80 degrees freezer in glycerol for later use in the measurements.<table id="partsT" style="width:100%"><tr><th>Construct</th><th>Promotor part from distribution</th></tr><tr><td>J23101-B0032-GFP</td><td>BBa_J23101</td></tr><tr> <td>J23105-B0032-GFP</td><td>BBa_J23105</td></tr><tr><td>J23106-B0032-GFP</td><td>BBa_J23106</td></tr><tr><td>J23115-B0032-GFP</td><td>BBa_K823005</td></tr><tr><td>J23116-B0032-GFP</td><td>BBa_J23116</td></tr><tr><td>J23117-B0032-GFP</td><td>BBa_J23117</td></tr><tr><td>J23101-B0034-BFP</td> <td>BBa_J23101</td></tr><tr><td>J23115-B0034-BFP</td><td>BBa_K823005</td></tr></table></p>';
document.getElementById("tab2").innerHTML = '<p>dfd</p>';
document.getElementById("tab2").innerHTML = '<p>dfd</p>';

Revision as of 18:03, 14 October 2014

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