Below is a general description of our services and approved SOM FY 2023/2024 fees.  Please contact Cheryl Bock for ES Cell and Embryo Services or Gary Kucera for Molecular Biology Services.  To download a request form click here.

ES Cell/Gene-Targeting Services

Generate HITI-Based Targeting Vector: $4297.00

Includes: Sequence verified, linearized, targeting vector DNA (Knockout, knockin, cKO, cKI model;) plus validated PCR assay and MOCK control DNA for screening  targeted ES cell clones.

 ES Cell Transfection of Targeting Vector: $4456.00

We have the best results with G4 ES cells which are a 129/B6N hybrid ES line we have obtained by MTA from Mount Sinai Hospital in Toronto.  Homozygous ES cells that are: 129S6/SvEvTac & also B6N ES cells can be used for transfection but this must be discussed before the targeting construct is made.

ES Cell Screening of Putative Targeted Clones: $1422.00

Up to 18 PCR screened positive ES cell clones will be processed and validated using PCR and sequencing.

 o    DNA Preparation only  from ES clones by RGEST for PI Lab: $594.00

Morulae Microinjection of Positive ES Cell clones: $4721.00

G4 ES cells of positive clones confirmed by the contracting lab or by the RGEST will be microinjected into ICR morulae to produce chimeric mice. Blastocysts from C57BL/6J will be injected for 129 or B6 ES cell lines. Blastocyst injection is a less efficient method to produce chimeric mice.

CRISPR/Cas9 Reagents and In-Vivo Genome Editing Services

Guide RNA design and cloning: $3766.00

Design and clone three independent guide RNAs into pX330 (or other) for one target locus.  Sequence Verify.

sgRNA Production via In-Vitro Transcription: $1512.00

Required for ALL CRISPR electroporation  requests, including Client-provided guide sequences.

RGEST prepares electroporation ready samples, No Exceptions!

Embryo Electroporation of CRISPR Reagents: $4456.00

PLEASE NOTE:  All KI models (point mutation, inserted sequence, etc.) require the RGEST to perform Founder Analysis.

Validated sgRNA (RGEST provided reagent only) + repair/donor oligo (if required) + cas9 protein.

 Founder Analysis, Initial: $1252.00

Genomic DNA preparation and sequence analysis of targeted locus from up to 24 founder animals to identify Indels. Founder Analysis by RGEST is a REQUIRED step for any KI project.

Founder analysis, Advanced, Allelic Subcloning: $2440.00

Subclone and sequence individual targeted alleles from up to 6 founders to identify/verify mutation

Pronuclear Injection of Client provided DNA for Transgenic Mouse Production Or BAC Transgene (RGEST or PI Lab)

DNA Microinjection to Produce Transgenic Mice: $4084.00

One-cell mouse embryos generated from a standard hybrid B6SJLF1/J cross, most efficient.

C57BL/6J embryos can also be used, but efficiency of transgenic production is much less than in hybrid background.  We provide NO guarantees that founders will be produced.

Special Trouble Shooting Assistance:

Labs are asked to confirm mutations in all of the chimeras from tail samples, and after breeding, in the offspring. If a lab encounters PCR difficulty, and needs our Core to help in troubleshooting, the service fee is $335. The Core does not offer a standard genotyping service. Contact the Breeding Core to explore this option.