Guidelines
CELL LINES
PLANT CELL LINES
PLANT VIRUSES
PLASMIDS
PHAGES
MICROORGANISMS
GENOMIC LIBRARIES
PROCEDURES
CATALOGUE
LABORATORY
PROCEDURES FOR PHAGES
REFERENCE NO.: PH/2003/01/01 TITLE: INTRODUCTION AND GENERAL DOCUMENTS Three resource centres have made their catalogues available via the CABRI server
namely, in alphabetical order:
The laboratory procedures described in these guidelines were derived from the
procedures used by these centres. They fulfill the requirements of the generally accepted
"Guidelines for the Establishment and Operation of Collections of Cultures of
Microorganisms" published by the World Federation for Culture Collections (WFCC) in
1999 (M/1999/1.00 Appendix 1) Each part of the guidelines consists of:
Procedure referencing is made according the following: Protocol code/Year/Chapter number/Procedure number
| Culture type | Protocol codes |
| Animal & Human cells | AHC |
| Bacteria, Archea, Yeasts, Fungi |
M |
| Phages |
PH |
| Plasmids | PP |
| Plant cells | PC |
| Plant viruses | PVC |
| Genomic libraries | GLI |
These guidelines describe basic procedures to be followed when new bacteriophages are accessioned and when stocks are quality checked whether they are master or distribution stocks. It does not contain working procedures for handling or isolation of bacteriophage DNA or RNA.
Bacteriophages are bacterial viruses that are not visible in a light microscope or easily centrifugable by other than high-speed laboratory centrifuging processes in order to recover pellets. Bacteriophages (= phages) are therefore only directly visible in an electron microscope and indirectly after lysis of their host cells, either in liquid culture due to clearing effects or on agar plates due to development of plaques. Principally, the lysogenic phages (integrated in the host chromosome) are to be distinguished from the free lytic phages (infective phage particles). Phages play an important role in teaching experiments, for the purpose of identification/classification/phage typing of bacteria, as model systems in modern virus research or as tracers in e.g. water quality laboratories.
Plaques are visible after lysis, they are zones in which the bacterial lawn has a lower growth density showing therefore a lower turbidity. They are characteristic for a particular phage. Phages are characterised in a number of ways. These include phage morphology as determined by electron microscopy, plaque morphology and type of nucleic acid present (single or double stranded DNA or RNA). Plaque morphology can vary from clear plaques produced by many virulent phage and hazy plaques produced by some lysogenic phage. Other phages produce plaques with clear centres and a hazy margin. All such descriptive properties contribute to characteristic descriptions of the various groups of phages.
RECOMMENDED PHAGE LITERATURE
Edward A. Birge: Bacterial and Bacteriophage Genetics, 3rd edition 1994, Springer New York Inc., ISBN 0-387-94270-X
Hans-Wolfgang Ackermann & Michael S. DuBow: Viruses of Prokaryotes, Vol. I and II, CRC Press Inc. 1987, ISBN 0-8493-6056-0 and 0-8493-6057-9
Joseph Sambrook & David W. Russell: Molecular Cloning, A Laboratory Manual, 3rd edition 2001, Cold Spring Harbor Laboratory Press, ISBN 0-879695773
Acknowledgement: We wish to thank our external reviewer Dr. Miguel Vicente, Centro Nacional de Biotecnologia, Consejo Superior de Investigaciones Cientificas (CSIC), Madrid, Spain, for his valuable comments on these Guidelines.
Guidelines
prepared for CABRI by DSMZ in cooperation with NCCB and NCIMB
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2023
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