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CRISPR/Cas9 – mediated gene targeting in mouse and human ES cells

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  1. Material modification service


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  2. Resource Description
    <b>IMPORTANT: To create economy of scale, at least 3 to 4 projects will be processed together as a batch order. Projects in a batch order may be from the same lab or from different labs. For each project at least 3 guide RNAs will be designed and constructed, unless otherwise specified.</b> Assuming 3 to 4 parallel projects (9-12 guide RNAs) in a batch order, the following prices apply: <b>Design and construction of gene specific Cas9-associated guide RNAs</b>: $300 per guide RNA <b>Design and construction of donor oligos or of large, complex targeting vectors:</b> Cost depends on type of donor DNA or targeting vector. Classical targeting vectors range from about $2,000 to $6,000. For donor DNA oligos, which are usually at least 150 nt long, total cost is the cost of synthesis at IDT plus a PGT processing fee and usually ranges from $200 to $300. Existing transgene constructs are accepted as basis for generation of targeting vectors, and prices will vary, depending on the length of the homologous arms to be added. <b>Transfection of one specific guide RNA with or without donor DNA into mouse ES cells</b> and generation of up to 45 ES cell clones by pre-selection with G418 or puromycin: $1,500. For an additional charge, pre-selection by GFP- FACS is also possible. The mouse ES cells can either be used to generate a mouse by blastocyst injection or just to measure effectiveness of a guide RNA. <b>Transfection of one specific guide RNA with or without donor DNA into human ES cells or human IPS cells</b> and generation of up to 45 clones by pre-selection with puromycin: $2,000. For an additional charge, pre-selection by GFP- FACS is also possible. <b>Genotyping by genomic PCR,</b> including detection of off-target effects, if necessary. For prices, please see the genotyping services section described under section A) above. <b>Karyotyping</b> for mouse ES cells is provided by the core as described under section A) above. Karyotyping of human ES cells is done for a flat charge of $700 per clone and includes growing the cells as well as G-banding of chromosomes.
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  4. Topic
    Homo sapiens
  5. Topic
    Mus musculus
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Copyright © 2016 by the President and Fellows of Harvard College
The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016