Biology Reference
In-Depth Information
Storage plate
Organize strains in an
intermediate storage plate
Intermediate
storage plate
Incubate overnight
at 37 °C
Replicate strains from the
frozen plate onto an agar
plate using a Boekel replicator
Agar plate
Incubate overnight
at 37 °C at 1050 rpm
Inoculate an overnight
culture plate using a
multichannel pipette
Use the overnight culture to
inoculate the experimental plate
FIGURE 1.2
Multiplexed manipulation of promoter fusion-containing strains for growth and expression
studies. Promoter fusion-containing strains of B. subtilis are individually picked from
transformation plates into a 96-well storage plate with a well capacity of 2 ml (plates with blue,
red, green, yellow spots). The outer wells of the plate are not utilized. Strains are grown
overnight at 37
C in these plates, glycerol is added and the plates are stored at
70
C.
To initiate expression analysis of the strains from a particular storage plate, a Boekel 96 pin
replicator is used to transfer an inoculum to agar medium on a square petri plate (blue arrow),
which is then grown overnight at 37
C. Should it be required to examine expression of strains
distributed among several storage plates, best results are obtained by generating an
intermediate storage plate fromwhich strains are then transferred to a square petri plate using
the Boekel replicator (red arrows). Overnight cultures of each strain are generated by
