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Staff:

Lucy Liaw, PhD
Core Director
(207) 885-8142
liawl@mmc.org

Anne Harrington
Core Manager
(207) 885-8189
harria@mmc.org


Transgenic Core Services and Fees

The Mouse Transgenic Core Facility provides a series of high quality services related to mouse genetics, manipulation of mouse embryos, re-derivation and cryopreservation, and the stocking and distribution of shared mouse strains. Questions regarding the availability of services should be addressed to Lucy Liaw, Ph.D., the Director of this resource (liawl@mmc.org).

 

 

Service

MMCRI investigator

Outside request

Pronuclear injection (FVB, other strains negotiable)

$400/day + mouse costs

$2800/line (guarantee of minimum 3 founder lines) + shipping costs. Other fees are associated with special requested strains.

Gene targeting

$1000 + mouse costs

Not accepted

Blastocyst injection

$800/line (minimum 2 days of injection) + mouse costs

$2800/clone + shipping

Cryopreservation

Variable, see description below

Not accepted

Regeneration of cryopreseved line

$200 + mouse costs

Inquires accepted

Re-derivation by oviduct transfer

$400/line + mouse costs

$2500/line + shipping

Note: It is required that the requesting investigator have an approved IACUC protocol before submission of a request to the Mouse Transgenic Facility.

*Any experiments involving the injection of cell lines provided by the requesting investigator require documentation of MAP testing indicating no detection of pathogens.

Pronuclear Injections for Transgenic Embryo/Strain Generation

The user submits a Pronuclear Injection Request Form and provides DNA for microinjection. When submitting DNA for pronuclear injection, please use this DNA purification protocol. As the quality of the DNA is critical for optimal success, extreme care must be taken in the preparation of the DNA, and the core will request documentation of the purity and quantitation of the purified sample.   Microinjection of DNA is performed routinely into FVB/N mouse embryos, although special arrangements can be made for other strains, with the requesting investigator covering additional mouse costs.  Injected eggs are transferred into a pseudopregnant recipient, and resultant pups are tagged and tail biopsied for the detection of the transgene by the investigator. Animals are housed within the core colony until genotyping is completed.  The founder animals are then transferred to the investigator.

Cost for pronuclear injections for in-house MMCRI investigators is $400.00/injection day.  Two days of injections are recommended for each construct to insure enough founders are produced. Approximately 100 eggs are injected each day.  In our experience, this leads to a minimum to three to up to 20 founder lines generated.

*The core facility is now accepting transgenic requests from investigators outside MMCRI for $2,800.00 per construct, yielding a minimum of three founder lines, assuming receipt of high quality DNA. Contact Lucy Liaw to make arrangements (liawl@mmc.org).

Embryonic Stem Cell Injections for Chimeric Mice

The investigator submits an ES Cell Injection Request Form and provides ES cells that have been tested and are negative for mycoplasma and mouse antibody production (MAP) against murine viral pathogens. ES cells are injected into C57BL/6 blastocysts.  Each investigator should submit at least 2 independent targeted clones to increase the chance of chimeric mice being produced.

The cost for ES cell injection for in-house MMCRI investigators is $800.00/clone. Two days of injections are recommended, with approximately 50 blastocyst injected.

Cryopreservation Program

The Transgenic Core is now offering embryo and sperm cryopreservation for your mouse lines. Please read this description of both procedures to help determine which option is best for you. Contact Anne Harrington with any questions (harria@mmc.org).

Embryo Cryopreservation

If you would like to maintain a line as a homozygote or your line is on an inbred background, this is the best option for you.

The line can be frozen two ways. First, you supply the core with 10 males that are fertile and capable of producing live offspring. Wildtype females will be ordered and used as egg donors to produce 2 cell embryos which will then be frozen. A sample straw will be thawed to verify viability. The Investigator is responsible for supplying proven young male studs, the embryo donors and covering per diem housing costs. The second option is to supply the core with 10 homozygous males and at least 30 (10/week) homozygous female egg donors. The females must be 3-4 weeks of age or 8-15 weeks of age. The per diem charge is applied.

Sperm Cryopreservation

This is the most cost effective option, however it has its limitations. The core will collect sperm from your males and freeze down 20 straws per male. Recovery of live mice from frozen mouse sperm is strain dependent. I recommend that you order an in vitro fertilization (IVF) service to test the sperm for their ability to generate viable pups. IVF results vary according to genetic background and the quality of individual males used for IVF. Thus we cannot offer a guarantee that any given IVF procedure will produce large number of pups.

Cryopreservation Costs

Embryo Freeze (one straw will be thawed and cultured to blastocyst stage)
The requesting investigator is responsible for mouse and per diem housing costs. $400.00/session

Embryo Recovery*
$100.00/recipient + per diem housing cost

Sperm Freeze
$100.00/male

In vitro fertilization
The requesting investigator is responsible for mouse and per diem housing costs.
$200.00/session
*vitrified embryos will not be accepted from MMRRC-UCD M

References:
Thornton C.E., Brown S.D.M, Glenister P.H. (1999). Large numbers of mice established by in vitro fertilization with cryopreserved spermatozoa: implications and applications for genetic resource banks, mutagenesis screens, and mouse backcrosses. Mammalian Genome 10: 987-992.

G.Charles Ostermeier, Michael V, Wiles, Jane S. Farley, Robert Taft (2008). Conserving, Distributing and Managing Genetically Modified Mouse Lines by Sperm Cryopreservation. PLos One 3:1-8

Re-derivation by Oviduct Transfer of Zygotes

The investigator submits a Re-derivation Request Form.  All mice that enter MMCRI from non-approved vendors MUST be re-derived and serology tested before housing within the barrier facility. 

The cost for re-derivation is $400.00/line.  Core staff will assist in determining requirement of mice for each line. The core will cover the cost of importation and re-derivation of all shared lines.



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