CRISPR and Molecular Biology Services
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) design and validation services and PCR Genotyping are also offered at the facility.
The Transgenic Facility provides an initial consultation to all investigators wishing to create a gene-modified mouse or cell line. After the consultation, reagents can be designed and validated. CRISPR reagent validation includes reagent design, reagent validation in embryos or a relevant cell line, and optimization of the PCR reaction for detecting mutations at the target site. For more information, refer to the Fee Schedule or send us an email at tgif@kumc.edu.
Responsibilities of the Investigator
- Provide the tail biopsies uniquely labeled and a positive control (either genomic DNA positive for the mutation or the plasmid used to generate the transgenic).
- Provide a proven PCR assay protocol and primers (100ul at 10mM concentration).
- Alternatively, provide the sequence of your target gene, and we will design a PCR reaction.
- Submit a Request Form in iLabs to initiate your project.
Services Provided by the Facility
- The facility will prep genomic DNA from tail biopsies using Qiagen's DNeasy Blood & Tissue Kit.
- The facility will select primers and optimize the PCR reaction conditions.
- The facility will perform a PCR for detecting the transgene(s) of interest.
- A report is provided to the investigator with gel documentation of the PCR results.
The facility can design and synthesize custom donor DNA for CRISPR projects. Contact Melissa Larson for more information.