1. Who can I contact for more information?
  2. What is the distribution fee for a cell line?
  3. After I order my cells, how long before I receive them (cells)?
  4. Once I know which cell lines I want, how do I order them?
  5. What is the parental cell line for the BayGenomics/SIGTR ES cells?
  6. What is the parental cell line for the Soriano (FHCRC) cells?
  7. What is resequencing?
  8. Have the cells I am ordering been pathogen tested?
  9. Where can I find more information on how to culture MMRRC ES cells?
  10. Is there any publication I can reference as to how the Gene Trap ES cell lines were made?
  11. What does the guaranteed microinjection service actually guarantee?
  12. How long between requesting ES cells and producing chimeras?
  13. What is the average germline transmission rate?
  14. How many chimeras do I need to breed to test for germline transmission?
  15. After producing chimeras, can you then test them for germline transmission?
  16. What is the cost of breeding chimeras?
  17. When can I expect germline transmission to be achieved?
  18. Why 12 weeks from when the chimeras are born?
  19. How long before my mice are shipped to my facility?
  20. How should I note the mice in publication, and acknowledge the donating party and MMRRC?
  21. What form of payment do you accept for these extended services?
 
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) 757-3290, sending a fax to (530) 757-3284, or sending an email to mmrrc@ucdavis.edu.
 
Additionally please review the MMRRC ES Cell Clones and Related Cell Lines page.
 
What is the distribution fee for a cell line?
For the distribution fees from the MMRRC, please see: http://www.mmrrc.org/catalog/fees.php.
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.
 
After I order my cells, how long before I receive them (cells)?
If resequencing is not requested, we anticipate the cells to be ready in about 4-6 weeks. If resequencing is requested,
we anticipate the cells to be ready in about 6-8 weeks. If the cell line cannot be verified on the first attempt, it may be
longer. Please keep in mind that we must receive your payment 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 payment.
 
Once I know which cell lines I want, how do I order them?
To place an order, please see instructions on this url: http://www.mmrrc.org/catalog/requestStocks.php
 
This includes a link to our catalogue search at this url: 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 service@mmrrc.org.
 
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 resequencing?
For an additional fee 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. If the sequence does not match, you will not pay for the cells or
the sequencing fee. As some cell lines do not confirm, we highly recommend that either you or we confirm the cell line
before you attempt to turn them into mice.
 
Please see our fees page for the cost at: http://www.mmrrc.org/distribution/fees.php, additionally please see our
protocol for Blasting the Sequence Tag to estimate the location of the trapping vector.
 
Have the cells I am ordering been pathogen tested?
Every ES cell line is tested for Mycoplasma spp. and Parvo spp. prior to shipment. The parental line 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.
 
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 clone protocols, please see the specific strain detail sheet for your cell line. Please search our catalog to find
the SDS for the MirKO cell line.
 
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.
 
What does the guaranteed microinjection service actually guarantee?
The guaranteed service guarantees at least one 50% male chimera is produced, or there will be no charge. This
guarantee does not extend to germline transmission.
 
How long between requesting ES cells and producing chimeras?
This can vary depending on additional services requested. Some cell lines do not sequence confirm, so we highly
recommend that either you or we confirm the cell line prior to injection. For ES cells, sequence verification, and
microinjection it can take 17-23 weeks until chimera pups are born.
 
What is the average germline transmission rate?
Over the last 4 years, the average germline transmission rate for chimeras derived by microinjection of gene trap ES cell
clones is about 76%.
 
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 fees for determining germline transmission are about $2500 per cell line. We will breed 2 male chimeras at a time.
The fee includes: per diems, colony management, female breeder purchase, genotyping litters, and housing positive
pups until they are 8 weeks of age. Please note this is only an estimate. The cost may be higher or lower depending on
the number of chimeras available, and their fertility and fecundity.
 
If additional breeding or housing is required beyond successful germline transmission, there will be additional fees.
Please contact us at mmrrc@ucdavis.edu for an estimate and more information.
 
When can I expect germline transmission to be achieved?
Please note that there is no guarantee for success. The earliest germline transmission can be determined is
approximately 12 weeks from when the chimeras are born.
 
We continue rotating females in with the males until they produce one or more hets, or 40 wild type pups. Once each
breeding chimera has produced at least 40 wild type pups or been non-productive for 90 days, we consider the project
failed. 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*
Tail snips 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 approx. 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?
“The mouse strain used for this research project, (insert strain name, see nomenclature rules or contact us for help)
was created from ES Cell clone (insert clone name here) obtained from the Mutant Mouse Regional Resource Center, a
NCRR-NIH funded strain repository, and was donated to the MMRRC by (the NHLBI supported BayGenomics gene trap
project, OR the Sanger Institute Gene Trap Resource, OR Philippe Soriano).”
Please contact us at mmrrc@ucdavis.edu for acknowledgement information on other MMRRC distributed ES cell clones.
Please note: The NIH has established a policy regarding Model Organism Sharing. We encourage you to contact us at
mmrrc@ucdavis.edu when you have published the strain so you can donate the mouse model back the MMRRC strain
repository for archiving and distribution.
 
What form of payment do you accept for these extended services?
For domestic customers we accept purchase orders (PO), credit card, check, and wire transfers. For international
customers we accept only credit card, check, and wire transfers. We do require payment prior to processing your
order. Please visit our payment information page for additional information and payment forms.