FAQ

 

 

What are the EuMMCR unit handling fee prices?

 

The EuMMCR unit price structure is the following:

targeting vectors (intermediate or final) 200 € ( ∼ 260 $)
one es cell line 500 € ( ∼ 650 $)
two es cell lines for one gene 700 € ( ∼ 900 $)
three es cell lines for one gene 900 € ( ∼ 1170 $)
four es cell lines for one gene 1400 € ( ∼ 1810 $)
five es cell lines for one gene 1900 € ( ∼ 2460 $)
development of genotyping reaction for chimeras,
using a short pcr strategy		 +200 € ( ∼ 260 $)
replacement or second vial of an es cell clone +200 € ( ∼ 260 $)

 

The handling fee does not include shipping. The EuMMCR unit requests from users their FedEx or (in Germany) TNT account numbers and uses this number for shipment.

 

How long does it take to receive knockout resources from the EuMMCR unit?

 

ES cells: Upon user request the EuMMCR unit thaws, expands and re-freezes several aliquots of the requested clone(s) in order to be able to provide the user with one aliquot and sustain the resource. Our controls include a PCR ensuring that the ES cell preparation is mycoplasma free and the verification or the genotype using one gene specific primer and one generic primer adjacent to the downstream loxP site. ES cell culture and the control means between 4-6 weeks lab work for us. From the lab's point of view, the EuMMCR unit is able to send the ES cell clones out after 4-6 week most of the times.


 

Targeting Vectors: Upon request the EuMMCR unit prepares DNA of targeting vectors, starting from the glycerol cultures we received from the EUCOMM vector production lab. For quality control we do restriction mapping on the vectors. This also will take between 4-6 weeks.

 

Are EUCOMM ES cells male or female?

 

EUCOMM ES cells are male.

 

Which wt ES lines have been used to generate conditional knockout clones?

 

wt ES cell line background Remarks
E14 129P2 Used for gene trap resource only. Widely used and well established. Suitable for injections on C57BL/6 blastocysts.
JM8.parental C57BL/6N GLT rate determined at the WTSI for JM8.parental ES cell clones: 62%
JM8.F6 C57BL/6N JM8 subclone isolated at the WTSI. The "F" stands for "feeder dependent". Suitable for injections in BALB/c or C57BL/6J-Tyrc/c blastocysts. GLT rate determined at the WTSI for JM8.F6 ES cell clones: 62%
JM8.N4 C57BL/6N JM8 subclone isolated at the WTSI. The "N" stands for "no feeders". Suitable for injections in BALB/c or C57BL/6J-Tyrc/c blastocysts. GLT rate determined at the WTSI for JM8.N4 ES cell clones: 69%
JM8A1.N3 C57BL/6N JM8.F6 derived ES cell line by removing the retroposon element in the Agouti gene by gene targeting. These cells are heterozygous for Agouti (A/a). This provides a convenient dominant coat color marker to detect germline transmission in testcrosses to C57BL/6N mice. JM8A1.N3 cells are feeder independent. With a (so far) small number the GLT rate of JM8A1.N3 clones determined: 75%

 

Is the Ordering of wt ES cells used in the EUCOMM program possible?

 

The ES cell lines JM8.F6, JM8.N4, and JM8A1.N3 are available from EUCOMM. Currently only the WTSI is distributing those wt ES cells. The the EuMMCR unit lab also has produced aliquots of JM8A1.N3 for distribution. However, these will be distributed as soon as these aliquots have been tested for germ line transmission (GLT).

 

What quality control procedures are implemented by EUCOMM/the EuMMCR unit to ensure non contaminated ES cell clones?

 

The EUCOMM production labs have MAP testing performed with wt ES cell stocks used in electroporations in order to generate ES cell clones. Each individual knockout clone is not MAP tested.

The EuMMCR unit lab tests every ES cell clone that is expanded in the lab for mycoplasma infection. So far not a single ES cell clone has been mycoplasma positive.

As a quality control effort EUCOMM is producing 320 mouse lines in five mouse clinics in Europe. One of these mouse clinics, which is currently receiving roughly 10% of the EuMMCR unit's ES cell production, has all imported ES cell clones MAP tested.

We view this quality control effort as batch testing of our ES cell production.

EUCOMM and the EuMMCR unit are committed to a very high standard of controls. However, EUCOMM and the EuMMCR unit cannot offer a "sanitary certificate" or a guaranty of any kind, since it is in the nature of the resource that the EuMMCR unit ES cells are handled in laboratories outside our supervision.