Abstract
To adapt pulsed-field gel electrophoresis (PFGE) to a busy clinical laboratory, we evaluated the use of achromopeptidase (ACP) as the sole agent for lysis of a variety of gram-positive cocci. Growth in an appropriate broth media was adjusted to a 3.0 McFarland density before aliquotting for washing and plug preparation. ACP incorporated into the agarose plugs, or added to the buffer solution, lysed cells in <1 h when suspended in a Tris-NaCl buffer. Two 30-rain washes in a Tris-EDTA buffer were sufficient to remove cellular debris without additional deproteination. Final PFGE results were obtained in -48 h, including the initial broth subculturing.
Original language | English (US) |
---|---|
Pages (from-to) | 288-291 |
Number of pages | 4 |
Journal | Diagnostic Molecular Pathology |
Volume | 6 |
Issue number | 5 |
DOIs | |
State | Published - Oct 1997 |
Externally published | Yes |
Keywords
- Achromopeptidase
- Molecular epidemiology
- Pulsed-field gel electrophoresis
ASJC Scopus subject areas
- Pathology and Forensic Medicine
- Molecular Biology
- Cell Biology