Team:IPN-UNAM-Mexico/Notebook/September

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Contents

01-September-2009

We do the transformation by head shock and plate.



02-September-2009

We haven't transformant cells.



07-September-2009

Meeting, we have to take new strategy for this ligation, J23100 does't work as backbone but as insert is so short.....the strategy is try again.

08-September-2009

We left overnight ligation 14ºC



09-September-2009

Transformation for head shock and plate. Get incubation overnight in 37ºC.



10-September-2009

We have only two colonies and have to analyze we picked out an get in LB amp agar liquid, then we put on the shacker in 37ºC.



11-September-2009

Do midi prep run out gel; it dosen't mark the correct weight.



14-September-2009

We going to use J23100 as backbone and Bba_K266003 as insert. (2385 bp)



21-September-2009

We going to change clormphenicol cassete Bba_P1010 for this we take off the ccdB gene and do restriction with P & E then ligate with Bba_K266006. Take out DNA from the well and electroporated the plasmid into Top 10 competent E. coli cells and plate, left overnight in 37ºC.



22-September-2009

We get colonies our ligation and left to incubate overnight on the shaker for midiprep.



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