inoculate the plate at 5 points with DH5α containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808032 BBa_K808032](L-arabinose regulated RBS-PhoA-His6tag-pNBEst13-Myctag-EstA) and incubate at 37°C
controll group is from the 6th point of same bacterial strain but carrying only [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808030 BBa_K808030] (RBS-PhoA-His6tag-pNBEst13-Myctag-EstA)
check every day for lysis area
LB-Agar-Bromthymol blue
stick in six sterile PET particles
inoculate the PET particles with DH5α containing pSB1C3-whole-construct and incubate at 37°C
check every day for colour change (pH decrease when PET gets digested)
controll group is on LB-Agar-Bromthymol blue without arabinose
DYT-medium
inoculate 50 mL DYT-medium-CAM with bacteria containing [http://partsregistry.org/wiki/index.php?title=Part:BBa_K808032 BBa_K808032]
incubate at 37°C to an OD600 = 0.6
incubate on ice for 15 min before inducing
prepare sterile test tubes
fill the tubes in the way shown in table 1
Component
tst tube 1
test tube 2
test tube 3
test tube 4
DYT-medium
x
x
x
x
PET particle
x
x
x
bacteria
x
x
x
induced
x
x
x
check every day the colour of the DYT-medium and take a picture
digestion of PET particles leads to a decreasing pH and will be shown by a yellow color
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