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p3 buffer from the amaxa p3 primary cell 4d-nuclefector x kit s  (Amaxa)

 
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    Amaxa p3 buffer from the amaxa p3 primary cell 4d-nuclefector x kit s
    P3 Buffer From The Amaxa P3 Primary Cell 4d Nuclefector X Kit S, supplied by Amaxa, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p3 buffer from the amaxa p3 primary cell 4d-nuclefector x kit s/product/Amaxa
    Average 90 stars, based on 1 article reviews
    p3 buffer from the amaxa p3 primary cell 4d-nuclefector x kit s - by Bioz Stars, 2026-03
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    Generation of a mouse model carrying a single-nucleotide deletion (C517) in exon 6 of the CYBB gene using CRISPR/Cas9-mediated HDR. ( A ) Scheme illustrating the HDR strategy for targeted deletion of C517 in exon 6 of the CYBB gene. Two guide RNAs (sgCybb#1 and sgCybb#2) were designed to target the desired region. Single-stranded DNA templates (ODNs) with a length of 125 nucleotides were synthesized as targeting donors for each sgRNA. The ODNs were designed to be homologous to the target region, except for the absence of a C at position 517. ( B ) To assess the activity of the guide RNAs (sgCybb#1 and sgCybb#2), the target region was amplified using the primers (Cybb_F2 and Cybb_R2), as indicated in ( A ). The resulting PCR products were incubated at 37 °C with crRNAs, tracrRNA, and recombinant SpCas9. The cleavage of the PCR products was analyzed by gel electrophoresis to evaluate the efficiency of the guide RNAs in inducing DNA cleavage. ( C ) <t>Electroporation</t> of ODN-Cybb#2 with sgCybb#2 and Cas9 into C57BL/6 mouse zygotes. Genotyping of the resulting founder mice was performed using PCR and Sanger sequencing with the indicated primers (Cybb_F1 and Cybb_R1). The genotypes of all founder mice are summarized in the table, with those carrying the desired genotype (∆C517), indicative of successful gene editing, highlighted in red. ( D ) Genotyping of F1 progeny derived from founder mutant #8327 by PCR (using indicated primers Cybb_F1 and Cybb_R1) and Sanger sequencing.
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    Generation of a mouse model carrying a single-nucleotide deletion (C517) in exon 6 of the CYBB gene using CRISPR/Cas9-mediated HDR. ( A ) Scheme illustrating the HDR strategy for targeted deletion of C517 in exon 6 of the CYBB gene. Two guide RNAs (sgCybb#1 and sgCybb#2) were designed to target the desired region. Single-stranded DNA templates (ODNs) with a length of 125 nucleotides were synthesized as targeting donors for each sgRNA. The ODNs were designed to be homologous to the target region, except for the absence of a C at position 517. ( B ) To assess the activity of the guide RNAs (sgCybb#1 and sgCybb#2), the target region was amplified using the primers (Cybb_F2 and Cybb_R2), as indicated in ( A ). The resulting PCR products were incubated at 37 °C with crRNAs, tracrRNA, and recombinant SpCas9. The cleavage of the PCR products was analyzed by gel electrophoresis to evaluate the efficiency of the guide RNAs in inducing DNA cleavage. ( C ) Electroporation of ODN-Cybb#2 with sgCybb#2 and Cas9 into C57BL/6 mouse zygotes. Genotyping of the resulting founder mice was performed using PCR and Sanger sequencing with the indicated primers (Cybb_F1 and Cybb_R1). The genotypes of all founder mice are summarized in the table, with those carrying the desired genotype (∆C517), indicative of successful gene editing, highlighted in red. ( D ) Genotyping of F1 progeny derived from founder mutant #8327 by PCR (using indicated primers Cybb_F1 and Cybb_R1) and Sanger sequencing.

    Journal: Genes

    Article Title: A Mouse Model of X-Linked Chronic Granulomatous Disease for the Development of CRISPR/Cas9 Gene Therapy

    doi: 10.3390/genes15060706

    Figure Lengend Snippet: Generation of a mouse model carrying a single-nucleotide deletion (C517) in exon 6 of the CYBB gene using CRISPR/Cas9-mediated HDR. ( A ) Scheme illustrating the HDR strategy for targeted deletion of C517 in exon 6 of the CYBB gene. Two guide RNAs (sgCybb#1 and sgCybb#2) were designed to target the desired region. Single-stranded DNA templates (ODNs) with a length of 125 nucleotides were synthesized as targeting donors for each sgRNA. The ODNs were designed to be homologous to the target region, except for the absence of a C at position 517. ( B ) To assess the activity of the guide RNAs (sgCybb#1 and sgCybb#2), the target region was amplified using the primers (Cybb_F2 and Cybb_R2), as indicated in ( A ). The resulting PCR products were incubated at 37 °C with crRNAs, tracrRNA, and recombinant SpCas9. The cleavage of the PCR products was analyzed by gel electrophoresis to evaluate the efficiency of the guide RNAs in inducing DNA cleavage. ( C ) Electroporation of ODN-Cybb#2 with sgCybb#2 and Cas9 into C57BL/6 mouse zygotes. Genotyping of the resulting founder mice was performed using PCR and Sanger sequencing with the indicated primers (Cybb_F1 and Cybb_R1). The genotypes of all founder mice are summarized in the table, with those carrying the desired genotype (∆C517), indicative of successful gene editing, highlighted in red. ( D ) Genotyping of F1 progeny derived from founder mutant #8327 by PCR (using indicated primers Cybb_F1 and Cybb_R1) and Sanger sequencing.

    Article Snippet: The cells were washed once with PBS at room temperature, and 3 × 10 5 cells were resuspended in 20 μL of AMAXA electroporation buffer.

    Techniques: CRISPR, Synthesized, Activity Assay, Amplification, Incubation, Recombinant, Nucleic Acid Electrophoresis, Electroporation, Sequencing, Derivative Assay, Mutagenesis