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  1. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ...positively transfected. Alternatively, you can use a selection marker to select transfected cells (for..., the most efficient and easiest way is to co-transfect several plasmid together, with each plasmid having...two spacers for each locus into PX330, then co-transfect or delivery the two plasmids together into your...run them and then select the best guide to co-transfect in HR donor. Alternatively, a new system described...oligo is fine based on new data). For general transfection, you could use 1ul of 10uM oligo stock for each... or mutation. Each arms is around 800bp. For transfection in cells like HEK, we use around 400ng of a ...single-point mutation, the best HR template for transfection is probably a single stranded DNA (ssDNA) oligo...

  2. CRISPR Plasmids - Drosophila

    Type
    Collection
    ... BspQI Transfection Puromycin yes, cut Ji-Long Liu 45946 pU6-BbsI-chiRNA dU6 BbsI Transfection none, need...Bullock and Port 51026 U6-BbsI-crRNA dU6 BbsI Transfection none, need Cas9 plasmid O'Connor-Giles, Harrison...

  3. Genomic Deletions in Mammalian Cell Lines

    Type
    Collection
    ...another transfection method such as cationic liposome-based transfection. Optimize transfection conditions...Perform maxi-prep for each CRISPR/Cas9 construct. Transfecting CRISPRs into Cells of Interest NOTE: This protocol...which is used for MEL cells. However, transient transfection of CRISPR/Cas9 plasmids may be successfully ...types using preferred culture conditions and transfection strategies for each cell type. While MEL cells...Fluorescence Activated Cell Sorting (FACS) of Transfected Cells Prepare cells for FACS by filtering them...

  4. TALEN Plasmids and Kits

    Type
    Collection
    ...Gate-assembled TALEN, both in microinjected embryos and transfected cells. TALEN expression is driven by the strong...expression in mammalian systems including hard-to-transfect cell lines. Features include (i) Esp3I (BsmBI)...on pTAL7a and pTAL7b for enrichment of double-transfected mammalian cells. In addition, a T7 promoter was...alternative. Based on transient double-selection of transfected cells, the system has successfully been employed...

  5. DNA Service - Cloning Grade DNA

    Type
    Collection
    ... different DNA concentrations. Can I directly transfect cloning grade DNA into cells? Cloning grade DNA...designed for use in cloning applications. Direct transfection into cells is not recommended. Can I order more...

  6. Plan Your Experiment

    Type
    Collection
    ...For easy-to-transfect cell types (e.g. HEK293 cells), transfection with standard transfection reagents may...or gRNA in a mammalian cell line that can be transfected at high efficiency Lentiviral transduction Cas9... mammalian cell lines Useful for difficult to transfect cell types, can also be used in vivo A common ...

  7. CRISPR Plasmids - C. elegans

    Type
    Collection
    ...(see paper) Transfection none, need Cas9 plasmid Calarco 47943 pMB70 cU6 BsaI Transfection none, need ...

  8. Luciferase Plasmid Collection

    Type
    Collection
    ...constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes... Renilla luciferase is used to monitor viral transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623... Firefly luciferase is used to moniter viral transfection Sidi Chen 109396 TBG-Vector Firefly TBG AAV-... Firefly luciferase is used to moniter viral transfection Sidi Chen 118412 ssAAV-EF1a-FLuc-WPRE-HgHpA_...

  9. Antibody Production

    Type
    Collection
    ...expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light...contain low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...antibody. Protein expression is compared to an untransfected control and the protein size is confirmed. Residual...

  11. Mammalian RNAi Tools

    Type
    Collection
    ...interfering RNA) via transfection, or shRNA (short hairpin RNA) via transfection or viral delivery of ...

  12. Zhang Lab CRISPR Page

    Type
    Collection
    ...human codon-optimized SpCas9; sgRNA can be co-transfected as PCR amplicons (Ran et al., Nature Protocols... #61422 - #61424 were designed for transient transfection : 61422 : Expresses dCAS9-VP64 activator with...codon-optimized SaCas9; SaCas9 sgRNA can be co-transfected as PCR amplicons using a protocol similar to ...

  13. Jaenisch Lab CRISPR Plasmids

    Type
    Collection
    ...dCas9 activators and sgRNAs can be separately transfected (pmax dCas9 activators [48223~48227] + sgRNA ...

  14. Adenovirus Plasmids

    Type
    Collection
    ...both plasmids. This DNA molecule can then be transfected into mammalian packaging cell lines to generate...

  15. Antibody Plasmid Collection

    Type
    Collection
    ...Collection . Once the plasmids are transiently transfected into mammalian tissue culture cells, the R-mAbs...

  16. Qi Lab CRISPR Page

    Type
    Collection
    ...selection or fluorescent gating of transiently transfected cells. Co-expression of both plasmids in HEK293...

  18. Brain Initiative Collection

    Type
    Collection
    ...flip-excision switch, for AAV production or direct transfection, slow variant Kaspar Podgorski 135421-AAV1 pAAV-syn-FLEX-axon-jYCaMP1s...flip-excision switch, for AAV production or direct transfection, slow variant Kaspar Podgorski 135424-AAV1 pAAV-syn-jYCaMP1s...Synapsin1 promoter, for AAV production or direct transfection, slow variant Kaspar Podgorski 135630-AAV1 pAAV-S5E2...

  19. Viral Vectors

    Type
    Collection
    ...the delivery of genetic information to hard-to-transfect mammalian cells. In addition, "viral pseudotyping...

  20. Viral Production

    Type
    Collection
    ... Vector Core (Link opens in a new window) . Transfections are performed using the transfer plasmid, a ...
Showing: 1 - 20 of 23 results