CRISPR/Cas9 services and off-target analysis
- gRNA design and production — Guide RNA (gRNA) will be designed and analyzed through CRISPRdirect and BLAST to determine potential off-target sites and analyzed with M-Fold to determine secondary structure.
- gRNA in vitro activity test — Transfection or electroporation of Cas9 and gRNA into appropriate cell lines will be done (along with repair template for knock-ins) and assessed with PCR, Cel-I, restriction enzyme digests, and/or sequencing as needed.
- Repair template design and production for knock-ins — Repair templates (ssODN, long ssDNA, or donor plasmids) will be generated with homology arms (when appropriate) to be used when making knock-in animals.
- Off-target analysis — Potential off-target sites will be determined via RGEN software tools and up to 10 potential off-target sites will be analyzed via PCR and DNA sequencing.
General molecular genetics and cloning services
Basic fee-based cloning services — General cloning services such as making plasmids for qPCR standards, TA cloning for sequencing of mixed PCR products, or complex cloning can be done on a contract basis.
Generation of targeting vectors/repair templates — Production of templates to generate conditional knock-out, knock-in, and transgenic animals can be purchased separately or as part of a larger contract to generate the animal model. These templates can be double-stranded plasmids (dsDNA), single-stranded oligonucleotides (ssODN), or long single-stranded templates (lssDNA).
Initial primer design and assay optimization available. DNA extractions from tail snips, ear punches or other tissue samples along with standard PCR or High Resolution Melt (HRM) analysis when appropriate. Digital droplet PCR (ddPCR) services also available for determining copy number of transgenes.
Organized and experienced management of rodent colonies is performed on an hourly rate. Services include setting up matings, weaning, tagging and snipping, and organized record keeping.
Long-term liquid nitrogen storage for embryonic stem (ES) cell lines and other samples.
A wide variety of services are available for animal model characterization. Services are customized to the needs of the investigator and include: histology, qPCR, imaging, behavioral analysis, and other phenotyping assays. Digital droplet PCR (ddPCR) services available to detect very subtle alterations in gene expression. Please contact us to set up a free consultation in order to determine how we can best assist you.
The AMC is available to discuss custom projects in which our molecular biology and animal modeling experience can be utilized. We have access to a wide range of equipment and have experience with many different techniques that may be beneficial to your research. Please contact us to set up a free consultation in order to determine how we can best assist you. The AMC can also partner on grant applications stating our ability to generate specific animal models or perform particular services.