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gene targeting
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gene targeting

The confluence of two groundbreaking technologies led to the first gene-targeted mouse models: plasticity of embryonic stem cells that contribute to the germline¹,² and successful homologous recombination³,⁴,⁵ between the ES cell genome and an introduced DNA sequence are typically known as a targeting vector. Gene targeting has been applied to understand mammalian physiology, to study human disease mechanisms, and pervaded biomedicine to create precision therapeutics.  NovoHelix provides a variety of options for engineering the desired genetic modification into your ES model. Our full-service projects include the entire ES cell model generation process from scoping out the design strategy and targeting vector construction to delivery of the mutant cell model. We also provide options to clients for subcontracting certain experimental workflows or phases of the project to us and many of these service descriptions are provided under our Support Services tab. A detailed cost estimate can be developed based on your specific project through our online Quote Request.
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Service

Catalog Nr

Service Description

Price

 Targeting Vector Construction
CMGT001

Construction of an isogenic gene targeting vector and Sanger-sequence verified with up to 20-kb insertion.  NovoHelix will provide an in silico map and provide restriction fingerprinting for overall plasmid structure via 3 digests. DNA is purified by a propriety AEX (anion exchange) chromatography and suitable for microinjection into mouse zygotes or nucleofection/electroporation.  The DNA is dialyzed on a membrane support in microinjection or electroporation buffer and then spun to remove particulate matter to obviate microinjection needle clogging.

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 ES Cell Gene Targeting (129P2/Ola ES Cells)
 CMGT002
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 ES Cell Gene Targeting (C57BL/6N ES Cells)
CMGT003
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 ES Cell Gene Targeting (BALB/cJ ES Cells)
CMGT004
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 ES cell 2nd Electroporation (Cre/Flp) (129P2/Ola ES Cells)
CMGT005
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 ES cell 2nd Electroporation (Cre/Flp) (C57BL/6N or BALB/cJ ES Cells)
 CMGT006

 Electroporation of a positive clone with recombinase (Cre or Flp), includes feeder preparation, passing picking, feeding, expansion of up to 12 clones, mycoplasma testing of up to 4 clones, preparing 3 clones for injections.

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 ES cell Screening by PCR and Southern Blot (NovoHelix-produced vector)
CMGT007

 Long-range PCR includes up to 300 clones; Southern blotting includes up to 18 samples with a validated probe (Neo, Hygro, GFP, Flpo, Cre)  Note: expansion of ES cells not included 

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 ES cell Screening by PCR and Southern Blot (Client-provided vector)
 CMGT008

 Long-range PCR includes up to 300 clones; Southern blotting includes up to 18 samples with a validated probe (Neo, Hygro, GFP, Flpo, Cre)  Note: expansion of ES cells not included 

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ES cell expansion and preparation for microinjection

(price includes up to 3 clones; client-provided ES cells without feeders )
CMGT009
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 ES cell expansion and preparation for microinjection

(price includes up to 3 clones, recommended for KOMP/EUCOMM cells; client-provided ES cells feeder-dependent )
 CMGT010
Expansion of an ES cell clone from one to many vials, including feeders if needed, freezing down, and mycoplasma testing. Service fee includes gamma-irradiated wildtype feeders.  Additional fees will be assessed for DR4 feeders and if selection is required.  NovoHelix has experience with FIAU, neomycin/G418, hygromycin, blasticidin, puromycin, zeocin and nourseothricin (clonNAT).
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