Difference between revisions of "Team:Exeter/Interlab"
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<p id="pp">Competent cells of E.coli DH5α were prepared following the provided protocol. Tested the OD of the cells using standard settings (see iGEM 2016 file),aiiming for a reading of 0.4-0.5. The first reading produced an average of 0.2, after 15 minutes the average increased to 2.5. The cells were spun down and re-suspended in TF-1 and TF-2 buffer. 100µl was aliquoted into 1 ml Eppendorf tubes and immediately dropped into liquid nitrogen. The cells were then left in the -80°C freezer. Chloramphenicol was made as a stock of 25mg/ml (weight measured was 76.6mg) and added to LB media to make plates for the iGEM interlab parts transformed with different constructs.</p> | <p id="pp">Competent cells of E.coli DH5α were prepared following the provided protocol. Tested the OD of the cells using standard settings (see iGEM 2016 file),aiiming for a reading of 0.4-0.5. The first reading produced an average of 0.2, after 15 minutes the average increased to 2.5. The cells were spun down and re-suspended in TF-1 and TF-2 buffer. 100µl was aliquoted into 1 ml Eppendorf tubes and immediately dropped into liquid nitrogen. The cells were then left in the -80°C freezer. Chloramphenicol was made as a stock of 25mg/ml (weight measured was 76.6mg) and added to LB media to make plates for the iGEM interlab parts transformed with different constructs.</p> | ||
| − | <p id="pp">The InterLab measurement kit contained 5 devices; (1) J23101+I13504, (2) J23106+I13504, (3) J23117+I13504, Positive and Negative control. The first attempt to transform 5µL of each device into our competent cells found no liquid in the tubes provided. The provided tubes were re-suspended using elution buffer from a Qiagen mini-prep kit. A transformation of plasmids used in the lab under a strong promoter and GFP was performed mirroring the | + | <p id="pp">The InterLab measurement kit contained 5 devices; (1) J23101+I13504, (2) J23106+I13504, (3) J23117+I13504, Positive and Negative control. The first attempt to transform 5µL of each device into our competent cells found no liquid in the tubes provided. The provided tubes were re-suspended using elution buffer from a Qiagen mini-prep kit. A transformation of plasmids used in the lab under a strong promoter and GFP was performed mirroring the Interlab protocol to determine if the cells were competent. Spread plates were made and incubated at 37°C.</p> |
<p id="pp">Competent cells containing the previously used strong promoter and GFP coding sequence showed strong expression of GFP. However the competent cells transformed with the iGEM parts showed no growth. A new interlab kit was ordered from iGEM.</p> | <p id="pp">Competent cells containing the previously used strong promoter and GFP coding sequence showed strong expression of GFP. However the competent cells transformed with the iGEM parts showed no growth. A new interlab kit was ordered from iGEM.</p> | ||
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<img src="https://static.igem.org/mediawiki/2016/1/1b/T--Exeter--interlabgraphforpart1egl.png" | <img src="https://static.igem.org/mediawiki/2016/1/1b/T--Exeter--interlabgraphforpart1egl.png" | ||
style="max-width:100%;margin:auto;display:block;"> | style="max-width:100%;margin:auto;display:block;"> | ||
| − | <span class="caption">Fig.</span> | + | <span class="caption">Fig. 3</span> |
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<img src="https://static.igem.org/mediawiki/2016/a/a6/T--Exeter--latinrectangleinterlabgraphforpart1egl.png" | <img src="https://static.igem.org/mediawiki/2016/a/a6/T--Exeter--latinrectangleinterlabgraphforpart1egl.png" | ||
style="max-width:100%;margin:auto;display:block;"> | style="max-width:100%;margin:auto;display:block;"> | ||
| − | <span class="caption">Fig.</span> | + | <span class="caption">Fig. 4</span> |
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Revision as of 13:09, 19 October 2016
