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Fees

Below is a general description of our services and fees.  Please contact Cheryl Bock for ES Cell and Embryo Services or Gary Kucera for Molecular Biology Services for further details.  To download a request form click here.

 

Services and Fees for transgenic model generation via homologous recombination in ES cells:

Full service KO, KI cKO, cKI model via Homologous Recombination: $11589

We will generate a targeting vector (KO,KI, cKO, cKI) via BAC Recombineering techniques, prepare the vector for ES cell targeting, generate a MOCK control DNA and validated PCR protocol for screening targeted ES cells. Targeting vector will be transfected into ES cells.  Clones will be selected and PCR screened using validated PCR protocol.  The BAC core will prepare genomic DNA from up to 18 putative positive ES clones and validate these clones across homology arms via a combination of PCR and/or Southern blot. Positive ES cell clones that have been validated will be injected into blasts, and then transferred to pseudopregnant females for generation of chimeric offspring.  Chimeras will be passed to the PI for further characterization/matings, etc.

 

Target vector generation:  $3601

We will generate a targeting vector (KO,KI, cKO, cKI) via BAC Recombineering techniques, prepare the vector for ES cell targeting, generate a MOCK control DNA and validated PCR protocol for screening targeted ES cells.

 

ES cell transfection with in-house generated targeting vector:  $3160

Targeting vector will be transfected into ES cells.  Clones will be selected and PCR screened using validated PCR protocol.  Up to 18 putative positive clones will be provided to the PI (or BAC core, see below) for further validation via Southern blot analysis.

 

ES cell clone validation (optional):  $1092

The BAC core will prepare genomic DNA from up to 18 putative positive ES clones and validate these clones across homology arms via a combination of PCR and/or Southern blot.

 

ES cell microinjection into blastocysts:  $3735

Positive ES cell clones that have been validated will be injected into blasts (C57BL/6NHsd), and then transferred to pseudopregnant females for generation of chimeric offspring.  Chimeras will be passed to the PI for further characterization/matings, etc.

 

ES cell microinjection with Repository clones provided by PI:  $2099

Repository clones provided by the PI lab (e.g. EUCOMM) will be injected into blasts (FVB/NHsd), and then transferred to pseudopregnant females for generation of chimeric offspring.  Chimeras will be passed to the PI for further characterization/matings, etc.

 

 

Services and Fees for transgenic model generation via CRISPR technology:

Full service CRISPR generated model (allelic analysis included):  $8846

We will generate up to three independent guide sequences and clone these into pX330 CRISPR vector. We will generate sgRNA from each of the three cloned guide sequences and screen for optimal activity using our in-vitro CRISPR screening protocol.  The sgRNA showing greatest activity will be used for pronuclear injections. The optimal sgRNA reagent and cas9 mRNA will be prepared and injected into the pronucleus of fertilized mouse embryos.  If no CRISPR effects (i.e. Indels) are identified (by CRISPR Founder screening), re-injections will be at no cost.  If indels are detected, but desired point mutations are not present (as determined by Allelic cloning(see below)), subsequent rounds of injection will incur additional charges at the original rate.  Genomic DNA will be prepared from tissue samples, followed by PCR amplification of the target region and Sanger sequencing. For further characterization of the indels, we will subclone the individual alleles from up to 6 founders.  These clones will be sequenced to identify the specific modifications to the target locus. 

 

CRISPR guide sequence Cloning:  $2507

We will generate up to three independent guide sequences and clone these into pX330 CRISPR vector.

 

CRISPR sgRNA Production via In-vitro Transcription:  $1140

We will generate sgRNA from each of the three cloned guide sequences and screen for optimal activity using our in-vitro CRISPR screening protocol.  The sgRNA showing greatest activity will be used for pronuclear injections.

 

CRISPR reagent Pronuclear Injection:  $2668

The optimal sgRNA reagent and cas9 mRNA will be prepared and injected into the pronucleus of fertilized mouse embryos.  If no CRISPR effects (i.e. Indels) are identified (by CRISPR Founder screening,) re-injections will be at no cost.  If indels are detected, but desired point mutations are not present (as determined by Allelic cloning(see below)), subsequent rounds of injection will incur additional charges at the original rate.

 

CRISPR Founder Screening:  $903

Genomic DNA will be prepared from tissue samples, followed by PCR amplification of the target region and Sanger sequencing.

 

Allelic cloning:  $1628

Note:  This service is required for requests involving base pair changes.

 For further characterization of the indels, we will subclone the individual alleles from up to 6 founders.  These clones will be sequenced to identify the specific modifications to the target locus.

 

Services and Fees for transgenic model generation via BAC Transgene:

BAC modifcations for Transgenics:  $2444

We will eliminate the single loxP site within the backbone vector of the BAC clone upon request.  We will modify the BAC clone per request (may include insertion of an ORF, reporter gene, etc).  BAC DNA will be maxi prepared and linearized for pronuclear injection.

 

Pronuclear Injection:  $2668

Linearized BAC DNA will be prepared and injected into the pronucleus of fertilized mouse embryos.   Additional injections, if required, will incur new charges at the original rate.