Application Note: Effective disruption of Pseudomonas aeruginosa biofilms using Adaptive Focused Acoustics (AFA)
In this technical note, we used the Illumina FFPE QC kit to assess the quality of DNA extracted from two different extraction technologies using four different types of FFPE samples.
The efficiency of several mechanical-based lysis and extraction techniques, such as Adaptive Focused Acoustics (AFA), probe sonication, and bead beating from yeast isolates was compared for (i) total protein yield, (ii) preservation of enzymatic activity, (iii) fragmentation of proteins, and (iv) protein bias (i.e., the failure to isolate specific proteins). Protein bias was determined from both the number and relative abundance of proteins separated by SDS PAGE and by two-dimensional gel electrophoresis (2-DE) analysis.
Adaptive Focused Acoustics (AFA) was used to effectively disperse Pseudomonas aeruginosa biofilms and disrupt bacterial cells in a low detergent concentration lysis buffer, significantly increasing the efficiency of protein extraction. Biofilm specific staining with Calcoflour White M2R revealed that overnight cultures of Pseudomonas aeruginosa PA14 produce extensive biofilms, and counterstaining with the cell permeable SYTO 62 red fluorescent nucleic acid stain indicate large numbers of intact cells encapsulated in the protective biofilm. Following a brief AFA treatment, Calcoflour White staining showed nearly complete disruption of biofilms along with a decrease in nucleic acid staining indicating lysis of over 94% of the biofilm encapsulated cells.
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