sacas9 mrna  (Millipore)


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    Name:
    Cas9 mRNA
    Description:
    Cas9 mRNA generated through in vitro transcription which has been capped and polyA tailed This mRNA should be used in conjunction with purified guideRNA The use of RNA vs plasmid constructs avoids complications with promoter embryo compatibility as well as the possibility of random integration of nuclease and gRNA expressing plasmids into the host animal genome
    Catalog Number:
    cas9mrna
    Price:
    None
    Applications:
    Functional Genomics/Transgenic Applications . Creation of gene knockouts in transgenic applications . Creation of knock-in animals with promoters, fusion tags or reporters integrated into endogenous genes
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    Structured Review

    Millipore sacas9 mrna
    Cas9 mRNA generated through in vitro transcription which has been capped and polyA tailed This mRNA should be used in conjunction with purified guideRNA The use of RNA vs plasmid constructs avoids complications with promoter embryo compatibility as well as the possibility of random integration of nuclease and gRNA expressing plasmids into the host animal genome
    https://www.bioz.com/result/sacas9 mrna/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sacas9 mrna - by Bioz Stars, 2020-05
    95/100 stars

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    Related Articles

    In Vitro:

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Synthesized:

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Mutagenesis:

    Article Title: Intelectin 3 is dispensable for resistance against a mycobacterial infection in zebrafish (Danio rerio)
    Article Snippet: .. 2000 pg of gRNA, 330 pg of cas9 mRNA (Sigma-Aldrich and Invitrogen, California, USA) and 1.5 ng of phenol red (Sigma-Aldrich) tracer were injected into one-cell-stage AB zebrafish embryos, and the success of mutagenesis was evaluated with the T7 endonuclease I (T7EI)- and the heteroduplex mobility assay (HMA) from isolated DNA of 2 dpf embryos . .. Gel images were obtained with ChemiDoc™ XRS+ system (Bio-Rad Laboratories) and analyzed with Image Lab software (v. 5.2; Bio-Rad Laboratories).

    Article Title: Intelectin 3 is dispensable for resistance against a mycobacterial infection in zebrafish (Danio rerio)
    Article Snippet: .. 2000 pg of gRNA, 330 pg of cas9 mRNA (Sigma-Aldrich and Invitrogen, California, USA) and 1.5 ng of phenol red (Sigma-Aldrich) tracer were injected into one-cell-stage AB zebrafish embryos, and the success of mutagenesis was evaluated with the T7 endonuclease I (T7EI)- and the heteroduplex mobility assay (HMA) from isolated DNA of 2 dpf embryos . .. Gel images were obtained with ChemiDoc™ XRS+ system (Bio-Rad Laboratories) and analyzed with Image Lab software (v. 5.2; Bio-Rad Laboratories).

    Isolation:

    Article Title: Intelectin 3 is dispensable for resistance against a mycobacterial infection in zebrafish (Danio rerio)
    Article Snippet: .. 2000 pg of gRNA, 330 pg of cas9 mRNA (Sigma-Aldrich and Invitrogen, California, USA) and 1.5 ng of phenol red (Sigma-Aldrich) tracer were injected into one-cell-stage AB zebrafish embryos, and the success of mutagenesis was evaluated with the T7 endonuclease I (T7EI)- and the heteroduplex mobility assay (HMA) from isolated DNA of 2 dpf embryos . .. Gel images were obtained with ChemiDoc™ XRS+ system (Bio-Rad Laboratories) and analyzed with Image Lab software (v. 5.2; Bio-Rad Laboratories).

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Article Title: Intelectin 3 is dispensable for resistance against a mycobacterial infection in zebrafish (Danio rerio)
    Article Snippet: .. 2000 pg of gRNA, 330 pg of cas9 mRNA (Sigma-Aldrich and Invitrogen, California, USA) and 1.5 ng of phenol red (Sigma-Aldrich) tracer were injected into one-cell-stage AB zebrafish embryos, and the success of mutagenesis was evaluated with the T7 endonuclease I (T7EI)- and the heteroduplex mobility assay (HMA) from isolated DNA of 2 dpf embryos . .. Gel images were obtained with ChemiDoc™ XRS+ system (Bio-Rad Laboratories) and analyzed with Image Lab software (v. 5.2; Bio-Rad Laboratories).

    Quantitative RT-PCR:

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Purification:

    Article Title: CRISPR/Cas9-Mediated Genome Editing and Mutagenesis of EcChi4 in Exopalaemon carinicauda
    Article Snippet: .. Preparation of Cas9 mRNA The pCMV-Cas9 vector (Sigma-Aldrich) was linearized by Xba I (Takara, Japan) and purified by ethanol precipitation. .. The purified linearized fragment was used as a template for in vitro transcription with a mMESSAGE mMACHINE T7 Ultra Kit (Ambion) to synthesize the capped and poly(A) tailed mRNA of Cas9.

    Expressing:

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Ethanol Precipitation:

    Article Title: CRISPR/Cas9-Mediated Genome Editing and Mutagenesis of EcChi4 in Exopalaemon carinicauda
    Article Snippet: .. Preparation of Cas9 mRNA The pCMV-Cas9 vector (Sigma-Aldrich) was linearized by Xba I (Takara, Japan) and purified by ethanol precipitation. .. The purified linearized fragment was used as a template for in vitro transcription with a mMESSAGE mMACHINE T7 Ultra Kit (Ambion) to synthesize the capped and poly(A) tailed mRNA of Cas9.

    Sequencing:

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Western Blot:

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Injection:

    Article Title: Intelectin 3 is dispensable for resistance against a mycobacterial infection in zebrafish (Danio rerio)
    Article Snippet: .. 2000 pg of gRNA, 330 pg of cas9 mRNA (Sigma-Aldrich and Invitrogen, California, USA) and 1.5 ng of phenol red (Sigma-Aldrich) tracer were injected into one-cell-stage AB zebrafish embryos, and the success of mutagenesis was evaluated with the T7 endonuclease I (T7EI)- and the heteroduplex mobility assay (HMA) from isolated DNA of 2 dpf embryos . .. Gel images were obtained with ChemiDoc™ XRS+ system (Bio-Rad Laboratories) and analyzed with Image Lab software (v. 5.2; Bio-Rad Laboratories).

    Article Title: Intelectin 3 is dispensable for resistance against a mycobacterial infection in zebrafish (Danio rerio)
    Article Snippet: .. 2000 pg of gRNA, 330 pg of cas9 mRNA (Sigma-Aldrich and Invitrogen, California, USA) and 1.5 ng of phenol red (Sigma-Aldrich) tracer were injected into one-cell-stage AB zebrafish embryos, and the success of mutagenesis was evaluated with the T7 endonuclease I (T7EI)- and the heteroduplex mobility assay (HMA) from isolated DNA of 2 dpf embryos . .. Gel images were obtained with ChemiDoc™ XRS+ system (Bio-Rad Laboratories) and analyzed with Image Lab software (v. 5.2; Bio-Rad Laboratories).

    Article Title: Utilising polymorphisms to achieve allele-specific genome editing in zebrafish
    Article Snippet: .. Microinjection Injection mixes were prepared by combining 1 μg of zebrafish codon optimised Cas9 mRNA ( ) with 500 ng of each gRNA and 0.5 μl Phenol Red (Sigma-Aldrich) in 2.5 μl total volume. .. 1 nl of each injection mix was injected into the yolk of 1–2-cell-stage embryos.

    Article Title: Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases
    Article Snippet: .. Cas9 mRNA and sgRNAs in M2 medium (Sigma-Aldrich) were injected into the cytoplasm of fertilized eggs with well-recognized pronuclei using a Piezo-driven micromanipulator (Prime Tech). .. The recombinant Cas9 protein: Foxn1 -sgRNA complex was diluted with DEPC-treated injection buffer (0.25 mM EDTA, 10 mM Tris at pH 7.4) and injected into male pronuclei using a TransferMan NK2 micromanipulator and a FemtoJet microinjector (Eppendorf).

    Article Title: CRISPR/Cas9-Mediated Deletion of Foxn1 in NOD/SCID/IL2rg−/− Mice Results in Severe Immunodeficiency
    Article Snippet: .. Cas9 mRNA (20 ng/μl) and sgRNA (20 ng/μl) were injected into the cytoplasm of fertilized eggs with visible pronuclei in M2 medium (Sigma) using a piezo-driven micromanipulator. .. The injected zygotes were cultured in potassium simplex optimized medium with amino acids at 37 °C under 5% CO2 in air until the blastocyst stage (3.5 d).

    Mouse Assay:

    Article Title: A non-inheritable maternal Cas9-based multiple-gene editing system in mice
    Article Snippet: .. Cas9 mRNA expression levels in Tg mice were measured by qRT-PCR using Cas9E-S and Cas9cla-A primers and NFCas9 RNA copy standard curves, as well as by CAG1620-S/Cas9215-A and Ramp3-S/Ramp3-A qRT-PCRs and western blotting with anti-FLAG; F1804, Sigma-Aldrich; St. Louis, MO, USA). mRNA copy number standard templates were prepared in a solution containing total RNA (50 ng) isolated from a wild-type 13.5-dpc embryo and serially diluted (10, 102 , 103 , 104 , and 105 copies) in vitro -synthesized Cas9 RNA (sequence length: 4146 nucleotides; 0.18 pg corresponding to 105 copies). ..

    Plasmid Preparation:

    Article Title: CRISPR/Cas9-Mediated Genome Editing and Mutagenesis of EcChi4 in Exopalaemon carinicauda
    Article Snippet: .. Preparation of Cas9 mRNA The pCMV-Cas9 vector (Sigma-Aldrich) was linearized by Xba I (Takara, Japan) and purified by ethanol precipitation. .. The purified linearized fragment was used as a template for in vitro transcription with a mMESSAGE mMACHINE T7 Ultra Kit (Ambion) to synthesize the capped and poly(A) tailed mRNA of Cas9.

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  • 95
    Millipore sacas9 mrna
    Sacas9 Mrna, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sacas9 mrna/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sacas9 mrna - by Bioz Stars, 2020-05
    95/100 stars
      Buy from Supplier

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