IGEM:Paris Bettencourt 2012/Notebooks/Semantic group/day by day//2012/09/20
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Result TransformationIt works weirdly, because there are colonies on positive control, none on negative control, and there is an absence of colonies in the middle of the ligation transformation plates, that can be explain by the platin on Cm on plain LBA plates, and we would have a gradient of antibiotic. So I took 2 colonies from each plate, the ones closer to the hole, and i make them grow in LB+Cm.The culture tube are composed of 5mL of LB + 5µL of Cm. I also made a PCR colony with the same colonies. Here is the content of 1 tube of PCR :
Here is the PCR pgm :
-> 25 cycles
Since the PCR is good, the sequence should be good too. We can register 2 new biobricks, and wait for the sequencing results
Amp 7*My only clone from yesterday grew up in LB+Amp, and I miniprep it and transform it in either NEB turbo or in MG1655 with or without the other plasmid that contain supD Result QCMI get 2 more colonies on Amp and 3 on Tet -> liquid culture. I also run a gel with the product of Digestion before transformation. The band corresponding to pSB1A1 can explain why we have colonies on Amp plates, but I still don't have colonies on Kan plates, and I have colonies on TetR, it might be contaminants here. Further experiments will make that clear. I will miniprep the liquid culture of this morning, and then transform them in either NEB or MG1655 with or without pSB1C3::supD. Preparation of TECAN experiment
I will to perform double transformation for this. |