IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/19: Difference between revisions
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! !! 260/280 !! ng/μL | |||
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| ECE 153 || 1.63 || 32.6 | |||
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| ECE 166 (1) || 1.73 || 56.2 | |||
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| ECE 166 (2) || 1.67 || 57.7 | |||
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| ECE 171 || 1.81 || 185.5 | |||
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==Transformation of IA771== | |||
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Revision as of 01:48, 20 August 2008
 Turing Pattern Formation
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Results of AgrA and C transformationTransformation has failed. No colonies were visible for the plates of Agr A or C. The Puc9 positive control grew. We believe that the problem was in the gel-extraction between ligation and transformation. Most of our plasmid was probably lost in this step. Next time we will directly use the results from the ligation reaction to transform. Although many bands were seen on the previous gel of the ligation reaction, we will check our transformation growth by single colony PCR to confirm transformation of the plasmid with our correct insert. Results from the transformation with glycerol stocksWe used two different glycerol stocks, one from 07/08 and one from 13/08. We also had 2 different protocols (cf 18/08 page). We tested our stocks on blank plates, cells are alive, the problem is to know if they are still competent or not.
- Stock from 07/08 : nothing on the control and nothing on the transformation plate. - Stock from 13/08 : nothing on the control and nothing on the transformation plate.
- Stock from 07/08 : nothing on the control and nothing on the transformation plate. - Stock from 13/08 : nothing on the control, and 1 colony on the transformed plate. We checked the fluorescence of bacteria into this colony (vector ECE166 with promoter and GFP). We have fluorescence, so our transformation is ok. The result of this confirm that our stock of competent cells in glycerol is still competnet since we managed to transform one colony. The problem is certainly not the storage, but our protocol when we use frozen competent cells> We need to improve the efficiency of this protocol. Test our Erythromycin stock- We put some colonies on a Ery 0.5 plate. They grow, but in a very small amount, so our antibiotic seems to be fine. - Cells in LB with antibiotic : no growth! So our antibiotic is ok, either we did not transform our cells and that is why everything grew on Ery plates, either there is a problem in our protocol for Ery test. Plasmid miniprep of ECE153, ECE166 and ECE171- Plasmid miniprep LB stocks from yesterday - Nanodrop
Transformation of IA771 | ||||||||||||||||
