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.
This Application Note describes the use of Covaris Adaptive Focused Acoustics (AFA) technology to extract DNA from FormalinFixed,
Paraffin Embedded (FFPE) tissues. When an AFA treatment is applied to a FFPE sample, it simultaneously deparaffinizes the
sample, emulsifies the paraffin, and rehydrates the tissue. This is a mechanical process which eliminates the use of hazardous organic
solvents. Using currently available methods, DNA extracted from FFPE tissues is not of sufficient quality for the sensitive, reproducible
molecular analysis required for clinical research and diagnostics. The Covaris FFPE DNA Purification Kit provides a high yield and high
quality solution for DNA extraction from FFPE tissue samples. Traditional quality control tests, such as DNA fragment size distribution
and concentration, are not indicative of the quality required for successful assays. Here, the quality of the extracted DNA with the
Covaris FFPE DNA Purification Kit is tested by a multiplex PCR-based quality control assay, showing the DNA is purified with yields
more than 3 times higher than other commercially available extraction methods and is more readily available for PCR amplification.
Overall, this new FFPE DNA extraction solution greatly enhances DNA availability, improves reproducibility, reduces hands on time,
and ensures the seamless integration with Next Generation Sequencing (NGS) and other molecular analysis applications, making it a
perfect solution for high value, often irreplaceable, clinical samples.
Covaris provides tools and technologies to improve pre-analytical sample preparation, enable novel drug formulations, and manage compounds in the drug discovery process.