P. larvae growth & Biofilm Pilot Study

Goal: Perform a rudimentary growth curve of P. larvae 9545 compared to several other bacteria strains. Also, determine the ability of P. larvae to form biofilms relative to closely related species. The growth curve will be performed using the continual plate reader located in the Pruess lab, and biofilm quantification will be performed using the crude crystal violet method.

Inoculated a single colony of bacteria into 5 mL of LB broth and incubated at 37C shaking at 225 rpm overnight. Each inoculation was performed in duplicate, resulting in two independent cultures for each of the six bacteria listed below.

Bacteria inoculated for study
P. larvae 9545
P. alvei 33A1
P. dendritiformus 30A1
B. cereus 6A5T
B. thuringiensis 4Q1
E. coli  B/r

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Friday October 31, 2014

Set up growth curve.

198 uL of LB broth was added to each well of a sterile 96 well plate. Added 2 uL overnight broth culture to each well, resulting in a 2:200 dilution (1:100). The cover was removed from the plate and a transparent adhesive cover was placed on top of it to prevent evaporation for the duration of the experiment. The plate was placed in the automatic plate reader in Pruess lab where it's optical density at 600 nm would be determined every hour for a total of 48 hours at room temperature would be calculated.

Plate set up:
There are two biological replicates of each strain, with eight technical replicates of each biological replicate.

Column 1 : P. dendritiformus

Column 2 :P. dendritiformus

Column 3 : P. alvei

Column 4 : P. alvei

Column 5 : P. larvae

Column 6 : P. larvae

Column 7 : B. cereus 

Column 8 : B. cereus 

Column 9 : B. thuringiensis

Column 10 : B. thuringiensis

Column 11 : E. coli

Column 12 : E. coli

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