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Escherichia coli NanoLuc®

BAA-2581-PACK

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A kit containing reporter-labeled Escherichia coli designed to distinguish control strain cross-contamination from true contamination in test samples. This product is appropriate for use as a positive control in quality control assays for E. coli.
Product category
Bacteria
Product type
Reporter-labeled bacterium
Type strain
No
Genome sequenced strain
Yes
Applications
Assay development
Food testing
High-throughput screening
Product format
Frozen
Storage conditions
-80°C or colder
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Price: $409.00 EA
Discounts may be available for our fellow nonprofit organizations.

Generally ships within 1-3 business days

Documentation

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.

Detailed product information

General

Specific applications
A kit containing reporter-labeled Escherichia coli designed to distinguish control strain cross-contamination from true contamination in test samples. This product is appropriate for use as a positive control in quality control assays for E. coli.

Food testing
Preceptrol
No

Characteristics

Serotype
O45:H2
Toxigenic
Yes
Toxin genes
eae (Intimin) positive
stx1 (Shiga toxin 1) positive
stx2 (Shiga toxin 2) negative
Comments
This strain was confirmed by PCR to carry the stx1 and eaeA genes (stx1+, stx2-, eaeA+).
Toxigenic

NanoLuc® and Nano-Glo® are registered trademarks of Promega Corporation. NanoLuc® and Nano-Glo® Technologies are licensed from Promega Corporation.

Handling information

Medium
Temperature
37°C
Atmosphere
Aerobic
Handling procedure
Kit Components (9 Reactions):
Component Format Volume Storage
Escherichia coli NanoLuc® Frozen in glycerol 3 vials  -80°C or colder 
Nano-Glo® Luciferase Assay Buffer Frozen in glycerol 490 µL  -20°C or colder 
Nano-Glo® Luciferase Assay Substrate Frozen in glycerol 12 µL  -20°C or colder 

Equipment and Materials Required but not Supplied:
Strain Propagation and Detection
Trypticase Soy Agar/Broth Inoculation loop
Water bath Eppendorf tubes
70% Ethanol Cotton swab
37°C incubator Laboratory paper (Whatman)
Pipette and tips Centrifuge

  1. Prepare a sterile test tube that contains #18 broth (4 to 5 mL).
  2. Thaw the sample in a 30°C to 35°C water bath, until just thawed (approximately 5 minutes). Immerse the ampoule just sufficient to cover the frozen material. Do not agitate the ampoule.
  3. Immediately after thawing, wipe down the ampoule with 70% ethanol and transfer the suspension into the broth tube. Mix well.
  4. Incubate the culture at 37°C for 24 hours.
  5. Use the culture as a control in assays for E. coli.

Detection of Control Strain
Swab Method:
  1. Prepare the reaction solution in an Eppendorf tube by mixing 1 µL of substrate with 49 µL of buffer.
  2. Collect several colonies with a cotton swab.
  3. Saturate the cotton swab with the prepared reaction solution.
  4. Visualize reporter expression in a dark area.
Paper Method:
  1. Prepare the reaction solution in an Eppendorf tube by mixing 1 µL of substrate with 49 µL of buffer.
  2. Saturate laboratory paper with the reaction solution.
  3. Collect several colonies onto an inoculation loop.
  4. Scratch colonies on the saturated paper.
  5. Visualize reporter expression in a dark area.
Broth Method:
  1. Prepare the reaction solution in an Eppendorf tube by mixing 1 µL of substrate with 49 µL of buffer.
  2. Centrifuge 500 µL of broth culture. Remove the supernatant.
  3. Resuspend the pellet in the reaction solution.
  4. Visualize reporter expression in a dark area.




Troubleshooting
Problem: The control strain does not luminesce after the addition of the reaction solution
Cause Solution
Over-passage of the strain, resulting in loss of the plasmid. Do not passage the strain more than 2 times.
The reaction solution is old or improperly prepared. Discard. Prepare reaction solution with ≥5% substrate and 95% buffer. Mix well.
Inadequate amount of reaction solution used. Ensure that your sample is fully saturated.
Contamination with another strain. Discard the culture. Begin with a fresh culture.

Problem: The frozen culture cannot be recovered
Cause Solution
Culture is not growing due to improper culture conditions. Check that the culture conditions (medium, temperature, and atmosphere) are appropriate. View culture periodically to check the condition. Increase time in culture after recovery.

Safety
See the appropriate Material Safety Data Sheets regarding safety precautions for the components of this product.
Handling notes

The Reporter-Labeled Control Strain Kit is stable at -80°C until the printed expiration date, provided there is no contamination. Care should be taken to minimize the number of freeze-thaw cycles when handling strains and reagents as it may affect the quality and shelf-life of the product. For best results, prepare fresh reaction solution by mixing the buffer and substrate before each use.  

This strain was confirmed by PCR to carry the stx1 and eaeA genes (stx1+, stx2-, eaeA+).
NanoLuc® and Nano-Glo® are registered trademarks of Promega Corporation. NanoLuc® and Nano-Glo® Technologies are licensed from Promega Corporation.
Additional information on this culture is available on the ATCC® web site at www.atcc.org.

Quality control specifications

Verification method
Whole-genome Sequencing

History

Depositors
J Lee, ATCC

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.

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.

This material is subject to the following restrictions in addition to those outlined in the ATCC Material Transfer Agreement:

  1. NanoLuc Label License 

For information on obtaining additional rights, please contact:

ATCC Licensing
Email: [email protected]

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

References

Frequently Asked Questions

Need assistance with this product? Contact our Technical Support team.

Telephone

Telephone

US and Puerto Rico

800-638-6597

Outside the US

+1-703-365-2700
hours

Hours of Operation

Monday-Friday
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