Team:Groningen/Labwork/9 August 2013

Claudio and Sander

Monitoring the grow of biofilm in liquid medium, LB agar and on titanium only.

Sebas

2 colonies on Pdes::cheY in munich backbone plate, did colony pcr the expected 675bp band was visible with some
aspecific bands. Inoculated both colonies in 3ml/LB/amp o/n at 37degrees celcius.
There were approx 150 colonies on the Munich+hyperspank+CFP plate. Inoculated 4col. in 3ml/LB/amp o/n 37degerees.
There were approx 100 colonies on the Pdes::cheY:eYFP in Munich backbone plate. All colonies were red indicating
that the P::RFP was still there.
Did a digestion on munich backbone with pstI and EcoRI with 1ug DNA for 30min at 37degrees celcius.
cut out the backbone from gel and stored at -20 degrees celcius
The 6 colonies for GFP and YFP in b.sub grew on new plates -> stabbed colonies on starch plate and incubated o/n
at 37degees celcius.

Inne & Sander

Received the motility knock outs today. Plated them on plates with various selection markers.

strain	description	selection marker
OI2838	cheB::cat	5 uL/mL cm
OI3305	cheC1 cheD1::cat	5 uL/mL cm
OI2737	cheW::cat	5 uL/mL cm
OI3017	cheB/cheR::cat	5 uL/mL cm
OI2652	delta cheR	5 uL/mL cm
OI2652	delta cheR	none
OI3305	delta cheC1	none
OI3061	cheW::cat,cheV::kan	5 uL/mL cm 10 uL/mL kan
OI3059	cheV::kan	10 uL/mL kan