Who can I contact for more information?
If you cannot find the answer to your question(s) here, you can always get more help by calling the MMRRC at UC Davis at (530) 757-3206, sending a fax to (530) 757-3284, or emailing firstname.lastname@example.org.
Additionally please review the MMRRC ES Cell Clones and Related Cell Lines page.
What is the distribution fee for a cell line?
Distribution fees vary based on the type of cell line requested; the fee includes expansion, pathogen screening, and genetic verification. For genetrap cell lines, genetic verification is resequencing. For all other cell lines, genetic verification includes a variety of tests that may include long-range PCR, qPCR, and copy number testing.
Fees for products and services are listed near the bottom of each Strain Detail Sheet in the catalog. Please note that there are additional handling and box fees. Once the order is placed online, an order estimate will be generated and emailed to you.
For shipping charges, we ask that you provide your own courier account number to cover the shipment of cells.
Can commercial, for-profit entities obtain MMRRC ES cells?
This varies depending on cell line. In general, only the KOMP targeted alleles are available to commercial entities.
After I order my cells, how long before I receive them (cells)?
For genetrap cell lines such as BayGenomics or SIGTR, we anticipate the cells will be ready in about 10-14 weeks. For all other cell lines such as MirKO or KOMP, we anticipate the cells will be ready in about 12-16 weeks. This timeline includes expansion, pathogen screening, and genetic confirmation. If the cell line cannot be verified on the first attempt, it may be longer. Please keep in mind that we must receive your billing information and an MTA or COU before we can begin processing your order.
The parental cell lines and the feeder cell lines are ready to ship out as soon as we receive the online order, MTA/COU, and billing information.
Once I know which cell lines I want, how do I order them?
To place an order, please see instructions at: http://www.mmrrc.org/catalog/requestStocks.php
You may access the MMRRC catalog search at: http://www.mmrrc.org/catalog/StrainCatalogSearchForm.php
If you need assistance with placing an order, please call the MMRRC customer service center at 1-800-910-2291 (North America) or 530-757-5710 (International). You may also email them at email@example.com.
What form of payment do you accept for these extended services?
For domestic customers, we accept purchase orders (PO), credit card, and checks. For international customers, we accept only credit card, check, and wire transfers. We do require billing information prior to processing your order. Please visit our payment information page for additional information and forms.
What is the parental cell line for the BayGenomics/SIGTR ES cells?
The parental cell line for BayGenomics and SIGTR ES cells is E14Tg2a.4, derived from mouse strain 129/P2OlaHsd. These can be ordered by requesting stock 015890-UCD.
What is the parental cell line for the Soriano (FHCRC) cells?
The parental cell line for these cells is AK7, which is 129S4/SvJaeSor derived. These can be ordered by requesting stock 015891-UCD. Feeder cells (SNL 76/7 STO) can be requested as well, stock 015892-UCD.
What is the parental cell line for the MirKO cells?
The parental cell line for the KECK MirKO ES cells is
E14Tg2a.4, derived from mouse strain 129/P2OlaHsd. These can be ordered by requesting stock 015890-UCD.
The parental cell line for the Sanger MirKO ES cells varies, please see the specific parental subline listed on each Strain Detail Sheet. There are two different parental cell lines that were used: JM8.F6 (050958-UCD) and JM8A3 (050960-UCD), both derived from mouse strain C57BL/6N.
What is the parental cell line for the KOMP cells?
The parental cell line for these cells varies, please see the specific parental subline listed on each Strain Detail Sheet. The JM8 and VGB6 parental lines are C57BL/6N derived.
There are six JM8 parental lines that are distributed by the MMRRC: JM8 (050957-UCD), JM8.F6 (050958-UCD), JM8.N4 (050959-UCD), JM8A3 (050960-UCD), JM8A3.N1 (050961-UCD), and JM8A1.N3 (050962-UCD).
The VGB6 parental line is not distibuted by the MMRRC. Please contact Velocigene to inquire about availability.
What is resequencing?
For genetrap cell lines such as BayGenomics or SIGTR, we perform 5’ RACE (Rapid Amplification of cDNA Ends) to check if the sequence we get matches that which is published by the donating institution. All mutant ES cell lines are heterozygous for the mutation, meaning they carry one modified allele and one wild type allele. If the sequence does not match, you will not pay for the cells.
Not all genetrap cell lines provide 100% gene knock out. Please check the location of the genetrap cassette within the gene; see our protocol for Blasting the Sequence Tag to estimate the location of the trapping vector.
How are knockout cell lines verified?
Genetic confirmation of the KOMP (CSD and Velocigene) and MirKO (Sanger and Keck) cell lines includes genetic verification at the 3′ and/or 5′ ends, vector copy number = 1, and presence or loss of Y chromosome. Type of genetic testing varies for CSD, Velocigene, MirKO targets. Approximately 30% of clones tested fail genetic analysis and will require a new clone be selected for expansion and testing. Based on the average germline rates, we recommend requesting and injecting 2-3 clones per targeted allele. All mutant ES cell lines are heterozygous for the mutation, meaning they carry one modified allele and one wild type allele.
Please note, there is high incidence of mixed cell populations in the KOMP-CSD targeted cell lines, and while our testing can detect this in most cases, it is not 100%, and there may be a very small percent of non or incorrectly targeted ES cells in the population, therefore we strongly recommend targeting verification of the derived mice with 5’ or 3’ LRPCR and/or Loss of Allele analysis.
We recommend performing chromosome counting or karyotyping prior to injection of clones (we offer chromosome counting as a added service).
What is chromosome counting?
Chromosome counting is performed to determine the percentage of euploid metaphase chromosomes in the targeted ES cells. A normal diploid mouse chromosome count is 40. We count at least 20 spreads. If 50% or more of the spreads counted total 40, the clone is considered to more likely contribute to both somatic cell chimerism and to germline transmission.
The fee for this service is $385.17 per clone for non-profit investigators.
Have the cells I am ordering been pathogen tested?
Every ES cell line is tested for Mycoplasma spp. and Murine Parvoviruses (MPV) spp. prior to shipment.
The parental lines for SIGTR and BayGenomics clones (E14Tg2a.4) and the parental line for Soriano clones (AK7.1) have had a full MAP test for which all pathogens were negative; please see the Health Status Report.
The parental sublines for KOMP have had IMPACT I testing for which all pathogens were negative; please see the respective health reports below for the specific KOMP parental subline.
- JM8 Health Status Report
- JM8.F6 Health Status Report
- JM8.N4 Health Status Report
- JM8A3 Health Status Report
- JM8A3.N1 Health Status Report
- JM8A1.N3 Health Status Report
We can perform additional screening for other potential pathogens upon request; please contact us for more information as the fee will be based on the test(s) requested.
Where can I find more information on how to culture MMRRC ES cells?
For culturing feeder-independent E14Tg2a.4 ES cells, including the BayGenomics and SIGTR collections, please see: http://www.mmrrc.org/strains/E14/Ctr_protocol.pdf
For ES cell lines derived from the Fred Hutchison Cancer Research Center, please see: Soriano Protocol
For MirKO and KOMP clone protocols, please see the specific strain detail sheet for your cell line. Please search our catalog to find the strain detail sheet for your cell line of interest.
For Lexicon Genetics Inc. donated cell lines, please see: Lex1 Protocol
For Izon donated cell lines, please see: Izon Protocol
For other protocols, please see:
- Soriano Protocol for X-gal Staining
- Protocol for Genotyping BayGenomics Mice
- SIGTR Protocols for Sanger Institute gene trap cell lines
- IGTC Protocols for some gene trap ES cell lines
Is there any publication I can reference as to how the Gene Trap ES cell lines were made?
Yes! Dr. William Skarnes has a publication in the following: Methods in Molecular Biology, Vol.97: Molecular Embryology: Methods and Protocols Edited by: P.T. Sharpe and I. Mason © Humana Press Inc., Totowa , NJ
Dr. Skarnes’ publication is Chapter 8 “Gene Trapping in Mouse Embryonic Stem Cells”.
Additionally we recommend the information available on the International Gene Trap Consortium (IGTC) website http://www.genetrap.org/ including the Gene trapping tutorial.
Is there any publication I can reference as to how the KOMP ES cell lines were made?
Yes! Methods used to create the CSD targeted alleles have been published, please reference the following: Skarnes, W.C., Rosen, B., West, A.P., Koutsourakis, M., Bushell, W., Iyer, V., Mujica, A.O., Thomas, M., Harrow, J., Cox, T. et al. (2011) A conditional knockout resource for the genome-wide study of mouse gene function. Nature, 474, 337-342.
Methods used to create the Velocigene targeted alleles have been published, please reference the following: Valenzuela, D. M., Murphy, A. J., Frendewey, D., Gale, N. W., Economides, A. N., Auerbach, W., Poueymirou, W. T., Adams, N. C., Rojas, J., Yasenchak, J., et al. (2003). High-throughput engineering of the mouse genome coupled with high-resolution expression analysis. Nat Biotechnol 21, 652-659.
How long between requesting ES cells and producing chimeras?
This can vary depending on the cell line and additional services requested. Some lines do not pass quality control testing, so would require selection of a new cell line or clone(s) for testing, which would add additional time to processing prior to shipment or microinjection. Microinjection requires an additional 11-15 weeks; this includes 2-6 weeks for scheduling and ordering donors, as well as aging of chimera to 6-7 weeks. Please note, production of chimera is not guaranteed.
For genetrap cell lines such as BayGenomics or SIGTR, testing of the ES cells and microinjection can take 21-29 weeks until chimera are ready to ship. This does not include additional time needed for chromosome counting (if requested).
For knockout cell lines such as MirKO or KOMP, testing of the ES cells and microinjection can take 23-31 weeks until chimera are ready to ship. This does not include additional time needed for chromosome counting (if requested).
What is the average germline transmission rate?
The average germline transmission rate for chimeras derived by microinjection of gene trap ES cell clones is about 76%.
The average germline transmission rate for chimeras derived by microinjection of KOMP ES cell clones is about 52%. Black (JM8) parental sublines are more germline efficient than agouti-corrected (JM8A) sublines. You can view germline rate by subline at https://www.komp.org/gltrates2.php.
How many chimeras do I need to breed to test for germline transmission?
We recommend breeding as many chimeras as possible once they are 6 weeks of age.
After producing chimeras, can you then test them for germline transmission?
Yes! Read below for even more information.
What is the cost of breeding chimeras?
The germline transmission testing fees are dependent on the parental cell line. Please contact us at firstname.lastname@example.org for an estimate and more information. Please note, germline transmission testing is not a guaranteed service.
When can I expect germline transmission to be achieved?
Please note that there is no guarantee for successful germline transmisison. The earliest germline transmission can be determined is approximately 12 weeks from when the chimeras are born.
Please view our Germline Transmission Testing Policy. If germline is not achieved, you are welcome to receive the original chimeras or we can discuss other options that may be available.
Why 12 weeks from when the chimeras are born?
- Chimerism assessed at 15 days of age (week 2)
- Chimeric pups weaned at week 3
- Chimeric males mated at week 6
- Litter born at week 9*
- Toe cuts for DNA extraction at week 10.5
- Genotyping and confirmation of germline transmission at week 12
*Please note that not all males mate immediately upon setting with females, and the mating date determines the dates of the latter steps.
How long before my mice are shipped to my facility?
We will contact you when chimeras are approximately 4 weeks of age to discuss shipping details. Health testing and report preparation takes about 2 weeks post weaning and submission. We must receive approval from your facility, confirmation of shipping address, and a courier account prior to shipment. The chimeras are usually 6-8 weeks of age at time of shipping.
How should I note the mice in publication, and acknowledge the donating party and MMRRC?
Near the top of the strain detail page of each cell line is a green “copy citation” button; click this button and paste the strain-specific citation where needed. This citation includes the strain name, RRID, repository, and donating party.
Please contact us at email@example.com if you have any questions about this citation.
Please note: The NIH has established a policy regarding Model Organism Sharing. We encourage you to donate the mouse model back the MMRRC repository for archiving and distribution. You can view more information about this process online.
Can I send mice that I made from MMRRC ES cells to another organization?
In general, MMRRC policy prevents an original recipient of MMRRC materials (e.g., ES cells, chimera, or germline-confirmed mice) to send or share that material with another (third party) investigator (“third party transfer”). Please see the MMRRC’s Third Party Transfers policy for more information.
How do I obtain assistance with biological, husbandry, breeding, or genotyping questions for a particular strain?
The MMRRC at UC Davis provides technical services for all products and services obtained from it. We can be contacted by emailing firstname.lastname@example.org or calling 1-530-754-6677.
How much are handling and shipping fees for ES cells?
In general, domestic shipments (U.S., not including Alaska, Hawaii, or insular territories) are subject to a $98.46 box and handling fee per ES Cell shipment and all international shipments (plus Alaska, Hawaii and insular territories) are subject to a $273.67 box and handling fee per ES Cell shipment. Special packaging requirements will incur extra fees. (Note: Mexico and Canada are international shipments requiring customs documentation.)
ES cells are shipped on dry ice. We request that customers provide a courier account number so that shipping can be billed directly to you by the courier. For domestic shipments, we use Federal Express (FedEx), and shipping fees are typically around $50-$150. We recommend using a specialized courier such as World Courier for international shipments, as they help with pre-approval of customs documentation and can re-fill dry ice if there are any delays/issues with customs once materials have shipped. Estimated international courier shipping fees are $1,600, but please contact the courier directly for a more accurate quote.
How much are handling and shipping fees for live mice?
In general, domestic shipments (U.S., not including Alaska, Hawaii, or insular territories) are subject to a $40 handling fee per live mouse shipment and $89.78 per crate used, and all international shipments (plus Alaska, Hawaii and insular territories) are subject to a $330.14 exportation fee (includes preparation of customs paperwork) and $179.56 crate fees per live mouse shipment. Special packaging requirements will incur extra fees. (Note: Mexico and Canada are international shipments requiring customs documentation.)
A specialized courier such as World Courier or Validated Courier is required for shipment of live animals. Estimated domestic courier shipping fees are $600 and international courier shipping fees are $1,600, but please contact the courier directly for a more accurate quote.