Team:Groningen/Labwork/20 August 2013
From 2013.igem.org
Claudio
BBa_J04450 is transformed to E. Coli DH5α on LB + Cm.This BioBrick has to aims: the part codes for an RFP, which is necessary for submitting new backbone to the registry and the backbone which must used to submit new part to the registry.
The synthetic DNA sequences are prepared for use. They are diluted in 20µl TE buffer.
Inne
The 0.7% agar plates that were to dry in the 37C incubator have dried. They were inoculated with liquid culture or from colonies on plates. The resulting in the following plates:# | strain | inoculated with |
1 | wt | toothpick from colony |
2 | wt | toothpick from colony |
3 | Delta cheY | toothpick from colony |
4 | Delta cheY | toothpick from colony |
5 | wt | 2,5 uL overnight culture |
6 | wt | 2,5 uL overnight culture |
7 | Delta cheY | 2,5 uL overnight culture |
8 | Delta cheY | 2,5 uL overnight culture |
9 | Negative control | empty toothpick |
New plates of 0.4% and 0.3% were prepared and left to dry at the bench. This to check the effect of different concentrations agar. For this the liquid cultures of cheY and the wildtype had to be put in new medium. The OD was measured of the existing cultures and new overnight cultures were made.
strain | selection marker |
wt | None |
delta cheY | cm (5 ug/uL) |
Sebas
Because the digestion check for the GFP didn't work new colonies were inoculated and the rest from the ligation wastransformed to E. coli (forgot heat shock at 37 degrees)