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Prices - updated Oct 1, 2016

The TMF currently offers the following services to investigators at UCI. Most services are also available to other UC campuses and to academic and commercial clients outside of the UC system. Please note - to be eligible for UC pricing, the funds must come from a UC account.

A note about strain nomenclature: C57BL/6NJ is the Jackson Laboratory substrain of C57BL/6N, and C57BL/6J is the Jackson Laboratory J substrain. These substrains are not genetically identical. B6SJLF1/J is a hybrid strain produced from crosses of C57BL/6J females with SJL/J males. When B6SJLF1/J female mice that have been mated with B6SJLF1/J stud male mice are used as egg donors, the fertilized eggs themselves (and hence the resulting transgenic founders) are referred to as B6SJLF2/J.

Production of Mice with Genomes Engineered by Cas9/gRNA

Services offered - see Cas9/gRNA Mediated Genome Engineering Services page for more information.

Design of Cas9/gRNA engineering - with client input, we will analyze the locus to be modified to identify target sites for Cas9/gRNA cutting. We will attempt to identify sites that have no predicted off-target sites on the same chromosome. We will design gRNA sequences as well as sequences of ssODN or plasmids with which to introduce the desired genome edit.

Ordering of commercial Alt-RTM gRNA's from IDT (recommended) - we will order Alt-RTM gRNA from IDT at IDT pricing, plus a modest administrative fee per gRNA. We routinely recommend clients use IDT Alt-RTM gRNA rather than use in vitro synthesized sgRNA's.

Synthesis of sgRNA - we will use in vitro transcription (IVT) to synthesize a single-guide (sg) RNA. Each IVT sgRNA is purified and quantified by gel electrophoresis.

Testing of cutting efficiency of gRNA / sgRNA - we will microinject gRNA or sgRNA with Cas9 mRNA and protein into fertilized mouse oocytes. The fertilized oocytes are cultured to blastocyst stage, then genomic DNA surrounding the targeted locus is amplified and analyzed using T7 endonuclease for evidence of cutting followed by NHEJ based repair. Two services are offered that differ based on the number of gRNA/sgRNA's tested.

Microinjection of Cas9/gRNA with HDR template

For cost of microinjection services associated with genome engineering via Cas9/gRNA, see the related information under DNA microinjection services.

Our Standard Microinjection service provides two independent days of microinjection, with transfer of surviving embryos to foster mothers. By performing the service on two different days we can minimize the likelihood of project delays due to factors out with our control. For example, despite ensuring the uterine morphology of pseudo-pregnant mice is favorable for transplantation of pre-implantation embryos at the time of embryo transfer, sometimes mice do not become pregnant. Despite minimizing noise and vibration in our vivarium, occasionally foster mothers cannibalize their offspring. Such events happen infrequently ~ 5 % of the time. However it is of little consolation if a client's project is delayed due to such events. By injecting on two independent days such delays become extremely rare. Most of our clients select the Standard (Two-Day) Microinjection Service and we are able to offer a reduced cost compared to ordering separate One-Day Services. However, if a client desires, we will accept requests to perform a one-day service.

Screening of pups for evidence of desired genome editing - we will obtain biopsy from pups between 8 - 10 days of age, extract genomic DNA and screen this using PCR based methods to identify evidence of introduction of desired genome edit. Please note, the price shown includes design of PCR primers and cost of restriction endonuclease, but is an estimate based on analysis of up to 20 pups. The real costs may increase if more than 20 pups are analyzed.

Ancillary services not described - we have experience analyzing the genomic changes produced in mice using Sanger Sequencing of cloned amplification products, as well as Sanger and Next-Gen Deep sequencing of amplification products from G0 generation mosaic animals. We can also design and validate PCR based genotyping assays, breed G0 animals and analyze genomes of G1 non-mosaic animals to verify introduction of the desired modification. For information and pricing on these and additional services, please contact Jon Neumann.

The information in the table below concerns pricing for services associated with Cas9/gRNA design and testing in mouse pre-implantation embryos.

Cas9/gRNA Genome Engineering Services

Service All UC clients* Non-UC clients** Commercial clients**
Design of Cas9/gRNA engineering strategy $850 $970 $1300
Ordering of IDT Alt-RTM gRNA Cost of gRNA from IDT, plus $10 administrative fee per gRNA. Cost of gRNA from IDT, plus $12 administrative fee per gRNA. Cost of gRNA from IDT, plus $16 administrative fee per gRNA.
Synthesis, purification and quantification of IVT produced sgRNA $220 $253 $345
Testing of cutting efficiency of gRNA / sgRNA (each of 3 RNA's injected into 20 fertilized oocytes) $900 $1040 $1380
Testing of cutting efficiency of gRNA / sgRNA (each of 6 RNA's injected into 20 fertilized oocytes) $1500 $1830 $2300
Screening of pups for evidence of desired genome editing (up to 20 pups) $740 $850 $1162

*Payment must be made from a UC account. Shipping fees are extra.

**Invoices will be mailed to the client. Credit card payments are not accepted. Shipping fees are extra.

Production of Transgenic Mice by Pronuclear Injection of DNA Constructs

More information is available about our DNA microinjection services.

Standard DNA microinjection service: DNA will be injected into fertilized eggs until at least 50 pups (or embryos) or at least 3 transgenic founder offspring have been produced, whichever comes first. To ID mice and provide a source of tissue for genotyping, the TMF will perform distal toe-biopsy on all offspring and deliver the tissue samples to the client for genotyping (via FedEx on dry ice for off-campus clients). Transgenic pups identified by the client are shipped or transferred after weaning. Transgenic status refers only to DNA integration. The TMF cannot guarantee expression of RNA or protein from the integrated DNA. Instructions on preparing the DNA are available. For plasmid-based transgenes, the final purification step will be performed by the TMF. To receive the service guarantee, all BAC vectors must be purified by TMF using transformed bacteria provided by the client. Brief descriptions of the construct and its expected phenotype are required.

Prices vary by donor strain and client’s affiliation, as described in the following table.

STANDARD DNA MICROINJECTION SERVICE
Donor strain: All UC clients* Non-UC clients** Commercial clients**
B6SJLF2/J $4424 $5100 $6800
Special Strain
$5773 $5773 $8900
C57BL/6NJ $4810 $5600 $7400

*Payment must be made from a UC account. Shipping fees are extra.
**Invoices will be mailed to the client. Credit card payments are not accepted. Shipping fees are extra.

One-Day DNA microinjection service: This service is only offered for B6SJLF2/J and C57BL/6NJ donors. If no transgenic founders are produced and the egg yield is more than one standard deviation below the average (based on the last 12 months), we will repeat the injection at no additional cost. Otherwise, the same conditions apply, as for our Standard service.

Prices vary by donor strain and client’s affiliation, as described in the following table.

ONE-DAY DNA MICROINJECTION SERVICE
Donor strain: All UC clients* Non-UC clients** Commercial clients**
B6SJLF2/J $2654 $3042 $4167
C57BL/6NJ $2623 $3006 $4118

*Payment must be made from a UC account. Shipping fees are extra.
**Invoices will be mailed to the client. Credit card payments are not accepted. Shipping fees are extra.

Production of Chimeric Mice by Injection of Targeted ES Cells into Blastocysts

More information is available about our ES cell injection service.

ES cell injection service: The client must supply at least one vial of frozen cells per clone. The TMF will thaw the vial, culture the cells through one passage, harvest blastocysts from C57BL/6J mice, and inject 10-15 ES cells into each viable blastocyst. Injected blastocysts will then be implanted into pseudopregnant females. The TMF guarantees that at least 40 blastocysts will be successfully injected and implanted into foster moms, or 3 chimeric pups will be produced, whichever comes first.

Testing ES cells for pathogens: ES cells supplied by the client or outside source (i.e., not produced by the TMF itself) must be tested for mycoplasma and mouse pathogens before injection. Clients must submit one extra vial of frozen cells for this purpose. Testing fee: $231/clone.

Expansion of ES cell clones to freeze back extra vials: This service is intended for cases where the client has obtained a single vial of cells, e.g., from KOMP or EUCOMM, and needs some back-up vials frozen down. The price varies according to the type of ES cells. Generally, 6-8 vials are frozen back, depending on the growth characteristics.

ES CELL INJECTION SERVICES
All UC clients* Non-UC clients** Commercial clients**
Injection (per day) $3433 $3947 $5390
Pathogen testing $231 $231 $231
Expand one vial to freeze back (feeder-independent cells) $553 $636 $868
Expand one vial to freeze back (feeder-dependent cells) $771 $887 $1210

*Payment must be made from a UC account. Shipping fees are extra.
**Invoices will be mailed to the client. Credit card payments are not accepted. Shipping fees are extra.

PRODUCTION AND CHARACTERIZATION OF TARGETED ES CELLS

More information is available about our gene targeting and Targeted Transgenesis services.

ES cell electroporation, clone picking, and DNA extraction: Using a targeting construct supplied by the client, we will electroporate the construct into embryonic stem (ES) cells, grow the cells under selection, pick up to 384 colonies, split the colonies into duplicate sets of plates, freeze one set, and extract DNA from the other set. To maximize the likelihood of success, we strongly urge clients to consult with us regarding the design of their targeting constructs and alternative methods of obtaining targeted ES cells before commencing their project.

Genotyping of ES cell colonies by Southern blot: We will screen the colonies from an ES cell electroporation to determine which ones have undergone homologous recombination. The client must supply a DNA probe that has been tested on a Southern blot of genomic DNA to prove it detects the correct wildtype band size. Note: the cost of restriction enzyme(s) used by the TMF is extra. The TMF also offers a service to identify and test probes for use in Southern analysis.

Expansion of selected ES cell colonies from 96-well plates: We will thaw colonies designated by the client, expand them, freeze down ~6-8 vials of cells from each colony, and extract at least 50 ug of DNA from each colony for further analysis of the targeted allele.

Chromosome counting: Cells are grown, metaphase spreads prepared, and digital images of spreads are analyzed to determine the number of chromosomes per spread, for a minimum of 20 spreads.

Secondary electroporation of targeted ES cell clones: We will perform a second electroporation on one targeted ES cell clone, to introduce an expression plasmid or similar construct. This service is intended for the transient introduction of a recombinase (Cre or Flp) to effect recombination of loxP or frt sites. Up to 192 colonies will be picked, split, frozen, and DNA extracted.

Targeted transgenesis: Transgene constructs with ROSA26 homology arms are electroporated into ES cells.  The cells are grown under selection and at least 24 colonies picked and split into duplicate plates.  One plate is frozen and DNA is extracted from the other plate.  The price includes Southern blot analysis of all picked colonies with a ROSA26 probe.

Small-scale Southern blot: This service can be used to test probes or to do a second Southern blot on selected ES cell clones, and can accommodate a maximum of 16 samples.  Cost of restriction enzymes is included. 

ES CELL CULTURE SERVICES
  All UC clients* Non-UC clients** Commercial clients**
Electroporation of feeder-independent ES cells $6041 $6947 $9484
Electroporation of feeder-dependent ES cells $7514 $8641 $11797
Southern blot analysis*** $3038 $3494 $4770
Colony expansion (see below) (see below) (see below)
Chromosome counting (price per clone) $400 $460 $628
Secondary electroporation of one feeder-independent ES cell clone $3600 $4140 $5652
Secondary electroporation of one feeder-dependent ES cell clone $4675 $5376 $7340
Electroporation to insert transgenes into ROSA26 locus $3600 $4140 $5652
Small-scale Southern blot*** $1102 $1267 $1730

*Payment must be made from a UC account.
**Invoices will be mailed to the client. Credit card payments are not accepted.
***Southern blot fees assume the client will supply probe DNA and tell us which restriction enzymes to use and what band sizes to expect.  An additional fee of $1102/probe will be added if the TMF is asked to design and test a probe.

Fees for expanding feeder-independent ES cell clones from 96-well plates*
# of clones expanded at one time: All UC clients Non-UC clients Commercial clients
 
1 clone $1336 $1536 $2098
2 clones $1576 $1812 $2474
3 clones $2525 $2904 $3964
4 clones $2886 $3319 $4531
5 clones $4182 $4809 $6566
6 clones $4462 $5131 $7005
7 clones $5384 $6192 $8453
8 clones $5758 $6622 $9040

*For feeder-dependent cells, add $168/clone.

MISCELLANEOUS SERVICES

More information is available about our breeding and cryopreservation/IVF/revival services.

Breeding Service: The TMF will breed genetically-modified mice in its holding rooms. The cost includes tail-cutting and identification of pups, with tail biopsies supplied to the client for genotyping. A flat fee per cage per day is charged, plus the cost of any mice purchased by the TMF.

Rederivation of mice by embryo transfer: The TMF will superovulate, mate, and collect preimplantation-stage embryos, using mice supplied by the investigator. The embryos will then be implanted into foster moms prepared by the TMF. Price includes serology of foster moms. This service is used for elimination of mouse pathogens and can accommodate a maximum of 30 females per session. We require at least 4 females per strain.

Cryopreservation of mouse embryos: Using both male and female mice supplied by the client, 8-cell stage embryos, harvested from superovulated and mated females, will be frozen and stored in liquid nitrogen in the TMF. The price is for one cryopreservation session, regardless of embryo yield. If sufficient embryos are obtained, the TMF will thaw a small portion and transfer them to foster moms to demonstrate ability to recover live mice. One session can accommodate up to 30 females. We require at least 15 females per strain.

Cryopreservation of mouse sperm: For each strain, the client must supply 2 male mice (we prefer proven breeders at 3-8 months of age). The TMF will collect sperm from the epididymis and vas deferens into cryoprotective media, aliquot the sperm into 20 semen straws, and freeze them in liquid nitrogen.  At least one extra straw will be frozen and thawed to test the sperm in an IVF assay, using oocytes from the same strain. Any strain producing fewer than 20% fertilized oocytes will be tested further by implanting fertilized embryos into foster moms. If live pups of the desired genotype are recovered we will consider the strain to be successfully preserved. Otherwise, we will repeat the freezing and IVF at no additional charge. Cost includes long-term storage in TMF liquid nitrogen tanks.

In vitro fertilization: Using mice supplied by the client, the TMF will harvest oocytes from superovulated females and fertilize them in vitro with either fresh or frozen sperm. Embryos are cultured overnight to the 2-cell stage and implanted into pseudopregnant foster moms. If no pups are obtained, the TMF will repeat the procedure once at no extra charge. Each service can accommodate up to 12 females.

PCR genotyping service: The TMF will perform PCR, using mouse genomic DNA, primers, and reaction conditions supplied by the investigator. The products will be separated by gel electrophoresis and photographed to document results. Each procedure can accommodate up to 24 DNA samples.

DNA isolation and genotyping service: The TMF will collect tail biopsies from mouse pups and ear-punch the pups for identification. DNA will be extracted from the biopsies and analyzed by PCR, using primers and a protocol of demonstrated utility, supplied by the investigator. Each procedure can accommodate up to 24 biopsies. This procedure is primarily intended for transgenic mice being bred by the TMF for an investigator.

MISCELLANEOUS SERVICES
All UC clients* Non-UC clients** Commercial clients**
Mouse breeding (per diem) $1.04/cage $1.32/cage $2.53/cage
Rederivation $1440 N/A N/A
Embryo cryopreservation (per session) $560 N/A N/A
Sperm cryopreservation (per strain) $510**** $630; $25/year storage $804; $37/year storage
Revival from frozen embryos (per strain) $480*** N/A N/A
IVF (up to 12 females) $1530 N/A N/A
PCR $115 N/A N/A
DNA isolation and genotyping $6.15/mouse N/A N/A

* Payment must be made from a UC account. Shipping fees are extra.
** Invoices will be mailed to the client. Credit card payments are not accepted. Shipping fees are extra.
***Pathogen testing of mice produced from frozen embryos is often necessary, especially if the embryos came from an outside source.  In such cases there will be an additional fee ($231) for the pathogen testing to test up to three samples.
****An administrative surcharge and storage fees will be billed for strains from other UC campuses.

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