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dcas9 protein  (OriGene)


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    Structured Review

    OriGene dcas9 protein
    Dcas9 Protein, supplied by OriGene, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/tp790148/Cas9+NC-NLS+Streptococcus+Pyogenes+Recombinant+Protein/pm37382191-342-56-58
    Average 91 stars, based on 4 article reviews
    dcas9 protein - by Bioz Stars, 2026-10
    91/100 stars

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    Related Articles

    CRISPR:

    Article Title: A Cost-Effective CRISPR/Cas9-Based Method for Sequencing the Functional Antibody Kappa Chain cDNA from Hybridomas Expressing Aberrant Kappa Chain mRNAs
    Article Snippet: Lastly, the cDNA was purified using Select-a-Size DNA Clean & Concentrator Kit (ZYMO Research, D4080). .. CRISPR/Cas9 digestion reaction was performed based on the following protocol: 3μl of the 10X CRISPR in vitro digestion buffer (20mM HEPES, 100mM NaCl, 5mM MgCl2, 0.1mM EDTA, pH 6.5), 1μl of 2μM sgRNA, 2μl of 1μM Cas9 protein, (Origene, TP790148), and 15ng of the cDNA (a molar ratio of 40:40:1) were added to a PCR tube and the volume was brought up to 30μl using nuclease free water. ..

    In Vitro:

    Article Title: A Cost-Effective CRISPR/Cas9-Based Method for Sequencing the Functional Antibody Kappa Chain cDNA from Hybridomas Expressing Aberrant Kappa Chain mRNAs
    Article Snippet: Lastly, the cDNA was purified using Select-a-Size DNA Clean & Concentrator Kit (ZYMO Research, D4080). .. CRISPR/Cas9 digestion reaction was performed based on the following protocol: 3μl of the 10X CRISPR in vitro digestion buffer (20mM HEPES, 100mM NaCl, 5mM MgCl2, 0.1mM EDTA, pH 6.5), 1μl of 2μM sgRNA, 2μl of 1μM Cas9 protein, (Origene, TP790148), and 15ng of the cDNA (a molar ratio of 40:40:1) were added to a PCR tube and the volume was brought up to 30μl using nuclease free water. ..

    Polymerase Chain Reaction:

    Article Title: A Cost-Effective CRISPR/Cas9-Based Method for Sequencing the Functional Antibody Kappa Chain cDNA from Hybridomas Expressing Aberrant Kappa Chain mRNAs
    Article Snippet: Lastly, the cDNA was purified using Select-a-Size DNA Clean & Concentrator Kit (ZYMO Research, D4080). .. CRISPR/Cas9 digestion reaction was performed based on the following protocol: 3μl of the 10X CRISPR in vitro digestion buffer (20mM HEPES, 100mM NaCl, 5mM MgCl2, 0.1mM EDTA, pH 6.5), 1μl of 2μM sgRNA, 2μl of 1μM Cas9 protein, (Origene, TP790148), and 15ng of the cDNA (a molar ratio of 40:40:1) were added to a PCR tube and the volume was brought up to 30μl using nuclease free water. ..



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    (A) shows the schematics of the proof of concept experiment. 4ug of the vector was first linearized using Seal (NEB) and rCutSmart buffer (NEB), the reaction is incubated for 1 hour at 37°C. The reaction then was run on a 1% agarose gel and purified. Next, the vector was digested using <t>CRISPR/Cas9</t> (3μl 10X Cas9 buffer, l00dng DNA, 1 μl of 2μM sgRNA, 2μl of lμM Cas9, nucleases free water to bring up the total reaction volume to 30μl) the reaction was incubated at 37°C for 60 min followed by 10 minutes at 65°C. (B) shows the reaction products and appropriate controls run on a 1% agarose gel at 100V for 30 minutes.
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    (A) shows the schematics of the proof of concept experiment. 4ug of the vector was first linearized using Seal (NEB) and rCutSmart buffer (NEB), the reaction is incubated for 1 hour at 37°C. The reaction then was run on a 1% agarose gel and purified. Next, the vector was digested using <t>CRISPR/Cas9</t> (3μl 10X Cas9 buffer, l00dng DNA, 1 μl of 2μM sgRNA, 2μl of lμM Cas9, nucleases free water to bring up the total reaction volume to 30μl) the reaction was incubated at 37°C for 60 min followed by 10 minutes at 65°C. (B) shows the reaction products and appropriate controls run on a 1% agarose gel at 100V for 30 minutes.
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    (A) shows the schematics of the proof of concept experiment. 4ug of the vector was first linearized using Seal (NEB) and rCutSmart buffer (NEB), the reaction is incubated for 1 hour at 37°C. The reaction then was run on a 1% agarose gel and purified. Next, the vector was digested using <t>CRISPR/Cas9</t> (3μl 10X Cas9 buffer, l00dng DNA, 1 μl of 2μM sgRNA, 2μl of lμM Cas9, nucleases free water to bring up the total reaction volume to 30μl) the reaction was incubated at 37°C for 60 min followed by 10 minutes at 65°C. (B) shows the reaction products and appropriate controls run on a 1% agarose gel at 100V for 30 minutes.
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    (A) shows the schematics of the proof of concept experiment. 4ug of the vector was first linearized using Seal (NEB) and rCutSmart buffer (NEB), the reaction is incubated for 1 hour at 37°C. The reaction then was run on a 1% agarose gel and purified. Next, the vector was digested using CRISPR/Cas9 (3μl 10X Cas9 buffer, l00dng DNA, 1 μl of 2μM sgRNA, 2μl of lμM Cas9, nucleases free water to bring up the total reaction volume to 30μl) the reaction was incubated at 37°C for 60 min followed by 10 minutes at 65°C. (B) shows the reaction products and appropriate controls run on a 1% agarose gel at 100V for 30 minutes.

    Journal: bioRxiv

    Article Title: A Cost-Effective CRISPR/Cas9-Based Method for Sequencing the Functional Antibody Kappa Chain cDNA from Hybridomas Expressing Aberrant Kappa Chain mRNAs

    doi: 10.1101/2023.06.08.544271

    Figure Lengend Snippet: (A) shows the schematics of the proof of concept experiment. 4ug of the vector was first linearized using Seal (NEB) and rCutSmart buffer (NEB), the reaction is incubated for 1 hour at 37°C. The reaction then was run on a 1% agarose gel and purified. Next, the vector was digested using CRISPR/Cas9 (3μl 10X Cas9 buffer, l00dng DNA, 1 μl of 2μM sgRNA, 2μl of lμM Cas9, nucleases free water to bring up the total reaction volume to 30μl) the reaction was incubated at 37°C for 60 min followed by 10 minutes at 65°C. (B) shows the reaction products and appropriate controls run on a 1% agarose gel at 100V for 30 minutes.

    Article Snippet: CRISPR/Cas9 digestion reaction was performed based on the following protocol: 3μl of the 10X CRISPR in vitro digestion buffer (20mM HEPES, 100mM NaCl, 5mM MgCl2, 0.1mM EDTA, pH 6.5), 1μl of 2μM sgRNA, 2μl of 1μM Cas9 protein, (Origene, TP790148), and 15ng of the cDNA (a molar ratio of 40:40:1) were added to a PCR tube and the volume was brought up to 30μl using nuclease free water.

    Techniques: Plasmid Preparation, Incubation, Agarose Gel Electrophoresis, Purification, CRISPR

    (A) shows the image of the CRISPR/Cas9 digested PCR product after the first digestion, the 560 bp band was purified and used as template for the second PCR. (B) shows the second PCR amplicon after the second CRISPR/Cas9 digestion. The 560 bp band was cloned and sequenced. (C) shows the sequencing results of the cloned fragment. This method decreased the percentage of the aberrant kappa chain transcripts by 88% in clone 262. (D) shows the comparison between method 1 versus method 2 in the percent reduction of the SP2/0 aberrant kappa chain sequences.

    Journal: bioRxiv

    Article Title: A Cost-Effective CRISPR/Cas9-Based Method for Sequencing the Functional Antibody Kappa Chain cDNA from Hybridomas Expressing Aberrant Kappa Chain mRNAs

    doi: 10.1101/2023.06.08.544271

    Figure Lengend Snippet: (A) shows the image of the CRISPR/Cas9 digested PCR product after the first digestion, the 560 bp band was purified and used as template for the second PCR. (B) shows the second PCR amplicon after the second CRISPR/Cas9 digestion. The 560 bp band was cloned and sequenced. (C) shows the sequencing results of the cloned fragment. This method decreased the percentage of the aberrant kappa chain transcripts by 88% in clone 262. (D) shows the comparison between method 1 versus method 2 in the percent reduction of the SP2/0 aberrant kappa chain sequences.

    Article Snippet: CRISPR/Cas9 digestion reaction was performed based on the following protocol: 3μl of the 10X CRISPR in vitro digestion buffer (20mM HEPES, 100mM NaCl, 5mM MgCl2, 0.1mM EDTA, pH 6.5), 1μl of 2μM sgRNA, 2μl of 1μM Cas9 protein, (Origene, TP790148), and 15ng of the cDNA (a molar ratio of 40:40:1) were added to a PCR tube and the volume was brought up to 30μl using nuclease free water.

    Techniques: CRISPR, Purification, Amplification, Clone Assay, Sequencing, Comparison