This service includes:

     - AAV-driven electroporation of C57BL/6J zygotes with engineered endonucleases.

    - Transgene-specific PCR to identify potential Founders

    - Long Read Sequencing of selected F1 mice (up to 3) to confirm proper integration.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

This service includes:

     - Pronuclear injection of C57BL/6J zygotes with engineered endonucleases.

    - Transgene-specific PCR to identify potential Founders

    - Long Read Sequencing of selected F1 mice (up to 3) to confirm proper integration.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

This service includes:

     - Small or large deletions in C57BL/6 zygotes. 

    - Gel electrophoresis of PCR amplicons (if applicable).

    - Sanger sequencing or targeted Next Generation Sequencing (NGS) to confirm proper deletion.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

This service includes:

     - Genetic modifications up to 200 nucleotides in C57BL/6J zygotes. 

     - Sanger sequencing and/or targeted Next Generation Sequencing (NGS) to confirm proper insertion.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

Injection of embryonic stem cells (ESC) or induced pluripotent stem cells (iPSC) into blastocysts to generate chimeric mice or study the contribution of cells to the developping embryos. A minimum of two recipient females will be implanted to generate chimeras.

Embryos will be frozen at the 2-cell stage. As part of our Quality Control (QC), the GEMF will thaw one straw of frozen embryos and culture them in vitro to the blastocyst stage to ensure future reviving of the embryos.

Frozen embryos will be implanted into recipient females to revive a Genetically Modified (GM) mouse line. A minimum of two recipient females will be implanted.

Expansion of ES cells generated outside of MDACC and cytogenetic analysis/Service Request form.

ES cell generation will include the isolation of the inner cell mass and subsequent growth of ES cells from blastocysts obtained from five to ten superovulated females.

Electroporation of investigator-provided construct into mouse embryonic stem cells/Service Request form plus documentation.

Policies:

Recombinant DNA Experiments

Because pronuclear injections and ES cell electroporations utilize unique DNA sequences that recombine into the mouse genome, the principal investigator needs to provide the GEM Facility with an approval number from the Institutional Biosafety Committee (IBC) prior to beginning these experiments.

Gene-Targeting in ES Cells

The investigator is responsible for demonstrating a reliable scheme for identifying targeted ES cell clones by Southern analysis with 5' and 3' probes by providing results from pilot experiments for this purpose. PCR analysis may be suitable for analysis if appropriate control experiments have been done. (Call Jan Parker-Thornburg for further information on certifying PCR analysis.) A convincing strategy for identifying mutant alleles must be demonstrated prior to the initiation of a gene-targeting experiment. 

n Vitro Fertilisation (IVF) will be performed from fresh, cold or frozen sperm to either generate live pups or to freeze embryos. A minimum of two recipient females will be implanted for reviving, otherwise the embryos will be frozen for archiving.

KI mice will be generated by AAV-driven electroporation of C57BL/6J zygotes using engineered endonucleases.

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

The donor plasmid used for Homologous Directed Repair (HDR) needs to be provided to the GEMF at high concentration for purification. Ideally, an endotoxin-free MidiPrep or a MaxiPrep should be provided to the GEMF.

KI mice will be generated by pronuclear injection of C57BL/6J zygotes using engineered endonucleases.

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

Small or large deletions will be performed in C57BL/6J zygotes. 

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

DNA extraction and Long Read Sequencing (LRS) for quality control of the editing events for up to three mice.

DNA extraction and Next Generation Sequencing (NGS) for quality control of the editing events.

DNA extraction and PCR genotyping to identify potential Founders.

A highly concentrated aliquot of the plasmid needs to be provided to the GEMF for purification. Ideally, an endotoxin-free MidiPrep or a MaxiPrep should be provided to the GEMF. Restriction enzyme(s) to isolate the linearised transgene also needs to be provided.

Transgenic mice will be generated by pronuclear injection of C57BL/6J zygotes.

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

Production of embryos and subsequent implantation into recipient females to eliminate pathogens from Genetically Modified (GM) mice. A minimum of two recipient females will be implanted to generate pups.

DNA extraction and Sanger sequencing to identify potential Founders.

Genetic modifications up to 200 nucleotides will be performed in C57BL/6J zygotes. 

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

Frozen sperm from two proven males will be frozen. As part of our quality control (QC), IVF will be performed. A project is considered complete if the fertilization rate equals or exceeds 20% for each male.

Electroporation of investigator-provided construct with 10-15 colonies picked for analysis/Service Request form.

Pricing is available upon request, after consultation with the investigator to determine the requirements of the specific project.

Frozen sperm from two proven males will be frozen. As part of our quality control (QC), IVF will be performed. A project is considered complete if the fertilization rate equals or exceeds 20% for each male.

Embryos will be frozen at the 2-cell stage. As part of our Quality Control (QC), the GEMF will thaw one straw of frozen embryos and culture them in vitro to the blastocyst stage to ensure future reviving of the embryos.

n Vitro Fertilisation (IVF) will be performed from fresh, cold or frozen sperm to either generate live pups or to freeze embryos. A minimum of two recipient females will be implanted for reviving, otherwise the embryos will be frozen for archiving.

Frozen embryos will be implanted into recipient females to revive a Genetically Modified (GM) mouse line. A minimum of two recipient females will be implanted.

Injection of embryonic stem cells (ESC) or induced pluripotent stem cells (iPSC) into blastocysts to generate chimeric mice or study the contribution of cells to the developping embryos. A minimum of two recipient females will be implanted to generate chimeras.

Production of embryos and subsequent implantation into recipient females to eliminate pathogens from Genetically Modified (GM) mice. A minimum of two recipient females will be implanted to generate pups.

A highly concentrated aliquot of the plasmid needs to be provided to the GEMF for purification. Ideally, an endotoxin-free MidiPrep or a MaxiPrep should be provided to the GEMF. Restriction enzyme(s) to isolate the linearised transgene also needs to be provided.

Transgenic mice will be generated by pronuclear injection of C57BL/6J zygotes.

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

Small or large deletions will be performed in C57BL/6J zygotes. 

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

KI mice will be generated by AAV-driven electroporation of C57BL/6J zygotes using engineered endonucleases.

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

The donor plasmid used for Homologous Directed Repair (HDR) needs to be provided to the GEMF at high concentration for purification. Ideally, an endotoxin-free MidiPrep or a MaxiPrep should be provided to the GEMF.

KI mice will be generated by pronuclear injection of C57BL/6J zygotes using engineered endonucleases.

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

Genetic modifications up to 200 nucleotides will be performed in C57BL/6J zygotes. 

Should another background be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

This service includes:

     - Small or large deletions in C57BL/6 zygotes. 

    - Gel electrophoresis of PCR amplicons (if applicable).

    - Sanger sequencing or targeted Next Generation Sequencing (NGS) to confirm proper deletion.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

This service includes:

     - Genetic modifications up to 200 nucleotides in C57BL/6J zygotes. 

     - Sanger sequencing and/or targeted Next Generation Sequencing (NGS) to confirm proper insertion.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

This service includes:

     - AAV-driven electroporation of C57BL/6J zygotes with engineered endonucleases.

    - Transgene-specific PCR to identify potential Founders

    - Long Read Sequencing of selected F1 mice (up to 3) to confirm proper integration.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

This service includes:

     - Pronuclear injection of C57BL/6J zygotes with engineered endonucleases.

    - Transgene-specific PCR to identify potential Founders

    - Long Read Sequencing of selected F1 mice (up to 3) to confirm proper integration.

Should a different background than C57BL/6J be required, additional costs will be added. Agistments costs passed the weaning date will be charged to the principal investigator.

DNA extraction and PCR genotyping to identify potential Founders.

DNA extraction and Sanger sequencing to identify potential Founders.

DNA extraction and Next Generation Sequencing (NGS) for quality control of the editing events.

DNA extraction and Long Read Sequencing (LRS) for quality control of the editing events for up to three mice.

Expansion of ES cells generated outside of MDACC and cytogenetic analysis/Service Request form.

ES cell generation will include the isolation of the inner cell mass and subsequent growth of ES cells from blastocysts obtained from five to ten superovulated females.

Electroporation of investigator-provided construct into mouse embryonic stem cells/Service Request form plus documentation.

Policies:

Recombinant DNA Experiments

Because pronuclear injections and ES cell electroporations utilize unique DNA sequences that recombine into the mouse genome, the principal investigator needs to provide the GEM Facility with an approval number from the Institutional Biosafety Committee (IBC) prior to beginning these experiments.

Gene-Targeting in ES Cells

The investigator is responsible for demonstrating a reliable scheme for identifying targeted ES cell clones by Southern analysis with 5' and 3' probes by providing results from pilot experiments for this purpose. PCR analysis may be suitable for analysis if appropriate control experiments have been done. (Call Jan Parker-Thornburg for further information on certifying PCR analysis.) A convincing strategy for identifying mutant alleles must be demonstrated prior to the initiation of a gene-targeting experiment. 

Electroporation of investigator-provided construct with 10-15 colonies picked for analysis/Service Request form.

Pricing is available upon request, after consultation with the investigator to determine the requirements of the specific project.