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Escherichia coli bacteriophage Lambda W60

97537

Product category
Viruses
Product type
Bacteriophage
Strain designation
LambdaW60 [W60]
Type strain
No
Product format
Freeze-dried
Shipping information
bacteria-free lysate
Storage conditions
2°C to 8°C
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Price: $414.00 EA
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Biosafety Icon BSL 1

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific applications
Potential bacteremia therapeutic agent
This is one of six bacteriophage strains deposited at the ATCC (ATCC® 97537™-97539™ are lambda strains and ATCC® 97540™-97542™ are P22 strains) that remain in the mouse circulatory system for long periods of time. They are potential therapeutic agents for bacteremias.
Preceptrol
No

Characteristics

Technical information
ATCC Technical Services does not have technical information on patent deposits that are not produced or characterized by ATCC. Additional information can be found in the corresponding patent available from the patent holder or with the U.S. and/or international patent office.

Handling information

Host
Escherichia coli MG1655 (ATCC 47076)
Medium
Temperature
37°C
Atmosphere
Aerobic
Handling procedure
  1. Follow general procedures given below for phage propagation.
  2. Escherichia coli strain MG1655 (ATCC 47076) is the recommended host.

GENERAL PROCEDURES FOR THE PROPAGATION OF BACTERIOPHAGE

To recover phage from freeze-dried or frozen vial:

  1. Prepare an actively growing culture of the recommended host strain before opening the phage specimen. The host should be 18-24 hours old. 
  2. Pick one colony from the isolation plate and homogenize in 5 mL of the appropriate broth. Incubate at 37°C while shaking until the growth reaches OD600 of 0.3 to 0.4. 
  3. Infect each 5 mL culture with 300 µL of the bacteriophage.  Add 25 µL each of 2M CaCl2 and 2M MgCl2.  Shake at 130 rpm at 37°C overnight. After 16 to 18 hours, centrifuge phage culture at 4000 g for 10 minutes. Filter the lysate with a 0.2 µm PES sterile filter then filter again with another 0.2 µm PES sterile filter.  The filtrate can be stored at 4°C.
  4. To perform a spot titer, warm one or two plates at 37°C. Overlay the surface with 2.5 mL of melted 0.5% agar (same medium) which contains 50 µL of an 18-24 hour host culture. The soft agar should be maintained at 43°C to 45°C until ready to pour. It may be advisable to use a water bath. Allow overlay to harden.
  5. The phage lysate can be serial diluted in a 96-well plate in quadruplicate (if desired). Aliquot 90 µL of broth medium (containing 25 µL each of CaCl2 and MgCl2 per 5 mL of broth) into each well. Add 10 µL of phage lysate to each well and mix. Pass 10 µL to each of the next set of wells and mix. Continue to the desired number of passages.
  6. Spot 2 µL of each dilution on the plate from step d. Up to 8 dilutions can fit on a 90 mm petri dish. After overnight incubation, lysis should be visible.  At the higher dilutions, individual plaques should be countable. pfu/mL = average plaque count / [(dilution factor) (2x10-3mL)]
    NOTE: Spotting the phage on plates makes visualizing the lysis easier. If phage is added directly to soft-agar before pouring plates, hazy or tiny plaques may be difficult to see. Resistant host bacteria may also mask plaque formation. 

To propagate phage: 
  1. Determine the total volume needed and place this amount of broth in a flask.  Add a small amount of overnight host culture to the flask and incubate at 37°C while shaking until the growth reaches OD600 of 0.3 to 0.4.   
  2. Infect with the calculated volume of phage lysate using the following formula. Volume of phage to add (ml) = (8x108 x total culture volume in ml x OD600 x MOI) / phage titer (PFU/ml). Add 25 µL each of 2M CaCl2 and 2M MgCl2 per 5 mL of broth.  Shake at 130 rpm at 37°C overnight.
  3. Centrifuge phage culture at 4000 g for 10 minutes. Filter the lysate with a 0.2 µm PES sterile filter then filter again with another 0.2 µm PES sterile filter.  The filtrate can be stored at 4°C.
    NOTE: 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. ATCC® uses double strength skim milk mixed half and half with the filtrate. Most phage can also be freeze-dried. ATCC® uses double strength skim milk mixed half and half with the filtrate.
Handling notes
The host strain (ATCC 47076) is a very robust culture. Only 50 µL are need per tube of soft agar.
Additional information on this culture is available on the ATCC® web site at www.atcc.org.

History

Depositors
National Institutes of Health/NIMH, CR Merril, National Institutes of Health/NIMH
Patent depository
This material was deposited with the ATCC Patent Depository to fulfill U.S. or international patent requirements. This material may not have been produced or characterized by ATCC.  As an International Depository Authority (IDA) for patent deposits, ATCC is required to complete viability testing only at time of initial deposit of patent material. Patent deposits are made available on behalf of the Depositor when the pertinent U.S. or international patent is issued, but material may not be used to infringe the patent claims.

Legal disclaimers

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.
Warranty

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.

Disclaimers

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. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Disclosures
This material is cited in a US and/or international patent and may not be used to infringe the claims. Depending on the wishes of the Depositor, ATCC may be required to inform the Depositor of the party to which the material was furnished.

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.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Curated Citations

Merril CR, et al. Long-circulating bacteriophage as antibacterial agents. Proc. Natl. Acad. Sci. USA 93: 3188-3192, 1996. PubMed: 8622911

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