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Staphylococcus aureus subsp. aureus bacteriophage 6


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Detailed product information



Handling information

Staphylococcus aureus subsp. aureus CDC 6 (ATCC 27704)
Handling procedure
  1.   Follow instructions below for titrating or propagating phage.
  2. The preferred method of propagation for this phage is either broth culture or soft-­agar overlay, using Staphylococcus aureus strain 6 (ATCC 27704) as propagation host.



To titrate phage:

  1. Prepare an actively growing broth culture of the recommended host strain before opening the phage specimen.  The host should be 18­-24 hours old.
  2. Pre­warm plates of the recommended medium in an incubator. Overlay the surface with 2.5 mL of melted 0.5% agar (same medium) that contains 0.1mL of the freshly grown host. The soft agar should be maintained 43 to 45°C until ready to pour. It may be advisable to use a water bath. Allow overlay to harden. 
  3. Add approximately 0.25 mL of the recommended broth to a freeze-­dried phage vial, 0.5 mL to a liquid cryovial.
  4. The rehydrated phage can be serially diluted by passing 0.25 mL of the phage into a tube containing 2.25 mL of the broth medium. Repeat for as many passages as desired. 
  5. 0.1 mL of each dilution is spotted on the surface of the prepared plates. Allow to dry. Three to four dilutions can be placed on each plate. After 18-24 hours incubation, lysis should be visible. At the higher dilutions, individual plaques should be countable.


To propagate phage (agar-overlay method):

  1. Phage may be propagated by preparing plates with the soft-agar/host overlay as above and covering the surface with approximately 0.5 mL of the concentrated phage.  Or, alternatively, you may add the phage directly to the melted agar/host before pouring over the plates.  For larger amounts, large-size T-flasks can be prepared with the recommended agar, and approximately 12.0 mL of melted soft-agar/host poured over the surface.  Phage is then allowed to run over hardened surface.  Phage may also be added directly to melted soft-agar before pouring as described above.
  2. After 18 to 24 hours incubation, the soft agar is scraped off the surface of the agar plates.  Centrifuge at about 1000 rpm for 25 minutes to sediment the cellular debris and agar.  Conserve the supernatant.
  3. This supernatant is passed through a .22 µm Millipore filter and the filtrate stored at 4-8°C.  Lysates should remain viable under refrigeration for long periods.  They may also be frozen with or without cryoprotectant.  If available, liquid nitrogen storage is the best method for long term storage.  Most phage can also be freeze-dried.  We use double-strength skim milk mixed half-and-half with the filtrate.
    NOTE:  Broth propagation methods may also be employed with most phage (see below).  Unless otherwise noted, ATCC uses the Adams agar-overlay method as described in M. H. Adams' Bacteriophages (Interscience Publishers, Inc., New York, 1959) for routine phage production.


For broth propagation:

  1. An overnight broth culture of the above host is aseptically transferred into a flask of the recommended broth so that a 1:100 dilution of the culture is achieved.
  2. To this, add the rehydrated phage.  Incubate for 6 hours with shaking.
  3. Harvest, centrifuge, and filter as described above.
Handling notes
Most staphylococcal phages will not produce marked clearing of a broth culture, and a somewhat turbid broth may or may not yield high-titer phage.  Phage resistant cells within the flask rapidly reproduce and form a turbid suspension concurrent with active phage liberation by sensitive cells.  It is therefore necessary to check the titer and repeat the process if insufficient phage have been produced.

Store filtrate at 4°C.  Storage at –20°C may cause inactivation of the phage. 

Additional information on this culture is available on the ATCC® web site at


Deposited as
AR Fodor

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Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.


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Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.


Frequently Asked Questions


Curated Citations

. Report of the subcommittee on phage-typing of Staphylococci of the international committee on nomenclature of bacteria, Moscow, July 1966. Int. J. Syst. Bacteriol. 17: 113-115, 1967.

. . Bull. World Health Organ. 24: 771-784, 1961.

Evins GM, Armfield AY. Effects of freezing on the stability of staphylococcal phages. Am. J. Clin. Pathol. 44: 480-481, 1965. PubMed: 4220988

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