Fees - Duke Cancer Institute Shared Resource
Below is a general description of our services and approved SOM FY 2019 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: $3,780
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: $3,800
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: $1,189
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: $500
Morulae Microinjection of Positive ES Cell clones: $4,000
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: $3,290
Design and clone three independent guide RNAs into pX330 (or other) for one target locus. Sequence Verify.
sgRNA Production via In-Vitro Transcription: $1,300
Required for ALL CRISPR electroporation requests, including Client-provided guide sequences.
RGEST prepares electroporation ready samples, No Exceptions!
Embryo Electroporation of CRISPR Reagents: $3,500
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: $1,000
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: $1,875.
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)
BAC Transgene Construction: $4,350
BAC Transgene construction, including removal of vector-specific LoxP site, Maxi prepared and linearized BAC DNA, analysis of final clone via pulse-field gel electrophoresis.
DNA Microinjection to Produce Transgenic Mice: $3,500
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.