Team:IPN-UNAM-Mexico/Notebook/September

=September=

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 J23100 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.

23-September-2009
We started the midi when program the centrifuge to 4000 rpm the falcon tubes broke down inside we lost the culture. Have to left inoculum again, we have lost this day.

24-September-2009
We start  midi - prep but whit 3500 rpm. We want to complete the midi. Completed the midi and carried out the gel.

25-September-2009
Next step digest L2-3 (BBa_K266000) as insert and attach to (BBa_K266006)