1352510631813_4b32bdfc1c708c768fff52926b47a7a3

Neurotransgenic Laboratory

Durham, North Carolina
1352510631813_4b32bdfc1c708c768fff52926b47a7a3
Neurotransgenic LaboratoryDuke UniversityDurham, North Carolina
  • 7 Services
  • 1 Member

The Neurotransgenic Laboratory (NTL) at Duke University conducts technology development research and provides services and training towards the genetic analysis of the nervous system.

The nervous system with its vast diversity of cell types and synaptic connections requires sophisticated tools to visualize and modify neuronal activity and interactions. Genetically engineered reporters and effectors allow such precise modifications. The Neurotransgenic Laboratory is focused on the design and generation of genetic tools for neuroscience research. Such tools include viruses for the precise stereotactical application of transgenes, plasmids and BACs for the generation of transgenic animals, genetically modified embryonic and induced pluripotent stem cells for differentiation and transplantation studies as well as for the generation of genetically modified animals.

The NTL offers coordinated expertise in the... Show more »

The Neurotransgenic Laboratory (NTL) at Duke University conducts technology development research and provides services and training towards the genetic analysis of the nervous system.

The nervous system with its vast diversity of cell types and synaptic connections requires sophisticated tools to visualize and modify neuronal activity and interactions. Genetically engineered reporters and effectors allow such precise modifications. The Neurotransgenic Laboratory is focused on the design and generation of genetic tools for neuroscience research. Such tools include viruses for the precise stereotactical application of transgenes, plasmids and BACs for the generation of transgenic animals, genetically modified embryonic and induced pluripotent stem cells for differentiation and transplantation studies as well as for the generation of genetically modified animals.

The NTL offers coordinated expertise in the generation of genetic tools for neuroscience research "from concept through creation", including experimental project design and planning, molecular construct preparation, high titer virus production, stem cell modifications, and mouse embryo microinjections.

Ultimately, the mission of the NTL is to provide an enriched environment for students, trainees, and established investigators and to develop, promote, and enhance the use of sophisticated genetic tools for understanding and resolving biomedical problems in basic and translational neuroscience research.

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    Adeno-associated virus (AAV) production
    $500.00 USD
    per Sample

    NTL provides services to investigators in the construction, purification, and titering of various viral vectors for research use.

    In addition, NTL provides consulting services for investigators who want to use virus vectors for laboratory and animal studies, and provides training and education on the handling of recombinant... Show more »

    NTL provides services to investigators in the construction, purification, and titering of various viral vectors for research use.

    In addition, NTL provides consulting services for investigators who want to use virus vectors for laboratory and animal studies, and provides training and education on the handling of recombinant viruses to interested researchers.

    Viral vector construction:

    starts at $500 for basic cloning (insertion of fragment into vector cassette); price increases with more complex assemblies.
    Generation of verified vector is guaranteed.

    Generating viral particles:

    $500 - 10 ml culture supernatant = 50 microliters concentrated stock
    $1,500 - 30 ml culture supernatant = 150 microliters concentrated stock
    Generation of virus stock is guaranteed; however, we cannot guarantee a specific titer. Average titers for concentrated stocks of small (promoter-reporter) lentiviral vectors are 10E11/ml, for larger cassettes 10E9/ml.

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    Blastocyst injection
    $3,500.00 USD
    per Sample

    lastocyst injections are carried out for generation of chimeric mice, i.e. mixtures of ES cell-derived and host blastocyst-derived tissues. The goal is a chimera with high contribution of ES cell-derived tissue, including the germline.

    ES cells for injection can be prepared by the investigator or by the NTL... Show more »

    lastocyst injections are carried out for generation of chimeric mice, i.e. mixtures of ES cell-derived and host blastocyst-derived tissues. The goal is a chimera with high contribution of ES cell-derived tissue, including the germline.

    ES cells for injection can be prepared by the investigator or by the NTL (preferred).

    Blastocysts (from strain C57BL/6 for 129-derived ES cells; from strain albino C57BL/6 for C57BL/6-derived ES cells) are injected with gene-modified ES cells and implanted into recipient dams. Chimeric males are turned over to the investigator at weaning age. We will supply a minimum of 3 chimeric males per project. While germline transmission is highly likely, we cannot guarantee it.

    129 ES cell lines: $3,500
    C57BL/6 ES cell lines: $4,500

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    ESC cell culture
    Embryonic stem cell culture
    $3,500.00 USD
    per Sample

    conventional targeting: $3,500/line
    ROSA locus targeting: $2,000
    Cre/Flp (cassette removal or cassette exchange): $1,500
    Generation of homologously targeted ES cell clones is guaranteed.

    conventional targeting: $3,500/line
    ROSA locus targeting: $2,000
    Cre/Flp (cassette removal or cassette exchange): $1,500
    Generation of homologously targeted ES cell clones is guaranteed.

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    Embryonic stem (ES) cell screening
    $3,500.00 USD
    per Sample

    The NTL carries out gene targeting experiments in embryonic stem (ES) cells. Routine targeting is done in strain 129-derived ES cell lines and in C57BL/6 ES cell lines.

    Genetic modifications are introduced in ES cells directly through homologous recombination of a targeting vector or through recombinase mediated cassette... Show more »

    The NTL carries out gene targeting experiments in embryonic stem (ES) cells. Routine targeting is done in strain 129-derived ES cell lines and in C57BL/6 ES cell lines.

    Genetic modifications are introduced in ES cells directly through homologous recombination of a targeting vector or through recombinase mediated cassette exchange (RMCE).

    Targeting vectors can be prepared by the investigator or by the NTL (preferred).

    Targeting constructs are electroporated into ES cells and selected for clones which took up the construct carrying the selectable marker. Clones are picked, expanded, aliquots are frozen and DNA is analyzed by PCR to identify clones with homologous recombination. Identified clones are expanded, and analyzed in detail by Southern blots before turning them over to the investigator or for injection into blastocysts.

    conventional targeting: $3,500/line
    ROSA locus targeting: $2,000
    Cre/Flp (cassette removal or cassette exchange): $1,500
    Generation of homologously targeted ES cell clones is guaranteed.

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    Lentivirus production
    $500.00 USD
    per Sample

    NTL provides services to investigators in the construction, purification, and titering of various viral vectors for research use. NTL routinely produces lentiviral vectors.

    Viral vector construction:

    starts at $500 for basic cloning (insertion of fragment into vector cassette); price increases with more complex... Show more »

    NTL provides services to investigators in the construction, purification, and titering of various viral vectors for research use. NTL routinely produces lentiviral vectors.

    Viral vector construction:

    starts at $500 for basic cloning (insertion of fragment into vector cassette); price increases with more complex assemblies.
    Generation of verified vector is guaranteed.

    Generating viral particles:

    $500 - 10 ml culture supernatant = 50 microliters concentrated stock
    $1,500 - 30 ml culture supernatant = 150 microliters concentrated stock
    Generation of virus stock is guaranteed; however, we cannot guarantee a specific titer. Average titers for concentrated stocks of small (promoter-reporter) lentiviral vectors are 10E11/ml, for larger cassettes 10E9/ml.

    « Show less
    Molecular cloning
    $500.00 USD
    per Sample

    The facility generates constructs for viral vectors, stem cell modifications, and transgenic and knockout mice. For the fastest generation of DNA constructs a combination of approaches are used: homologous recombination in bacteria (recombineering), PCR stiching, DNA synthesis, Choo-Choo cloning, as well as traditional cloning using... Show more »

    The facility generates constructs for viral vectors, stem cell modifications, and transgenic and knockout mice. For the fastest generation of DNA constructs a combination of approaches are used: homologous recombination in bacteria (recombineering), PCR stiching, DNA synthesis, Choo-Choo cloning, as well as traditional cloning using restriction enzymes and DNA ligases.

    The NTL carries out de novo construction of vectors for standard marker expression, overexpression and dominant-negative expression, gene-swap, and gene replacement, as well as of vectors incorporating conditional and inducible mutagenesis strategies for cell-specific and temporal-specific expression and deletion using Cre and Flp recombination systems and Tet-on/off combinations, respectively.

    Constructs carrying the desired genomic modifications can be used for viral delivery of the transgene, for cell modifications, and to generate transgenic mice which will carry the transgene at a random integration site in the genome or to generate mice which will have exchanged the endogenous locus with the construct via homologous recombination in embryonic stem cells.

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    Transgenic mouse
    Transgenic mouse development
    $3,000.00 USD
    per Strain

    Zygote injections are carried out for generation of transgenic mice, i.e. mice carrying the transgene randomly integrated into the genome. DNA is injected into the male pronucleus. DNA for injection can be plasmid inserts or linear BAC DNA.

    DNA can be prepared by the investigator or by the NTL (preferred).

    Zygotes can... Show more »

    Zygote injections are carried out for generation of transgenic mice, i.e. mice carrying the transgene randomly integrated into the genome. DNA is injected into the male pronucleus. DNA for injection can be plasmid inserts or linear BAC DNA.

    DNA can be prepared by the investigator or by the NTL (preferred).

    Zygotes can be injected from hybrid strains (B6SJLF1) or from inbred strains (C57BL/6).

    Injected zygotes are implanted into recipient dams. Pups are genotyped by NTL; identified founders are turned over to the investigator at weaning age (21 days). If DNA for microinjection is prepared by NTL, we will guarantee a minimum of 3 founders.

    F1 hybrid strain: $3,000
    C57BL/6 inbred strain: $4,000
    Delivery of 3 founders is guaranteed for both plasmids and BACs (expression of transgene cannot be guaranteed).

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