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smai restriction enzyme  (New England Biolabs)


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    New England Biolabs smai restriction enzyme
    Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of <t>extracted</t> <t>rAAV</t> genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with <t>SmaI.</t> (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).
    Smai Restriction Enzyme, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 2723 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/smai/SmaI/pmc13049938-281-0-5
    Average 97 stars, based on 2723 article reviews
    smai restriction enzyme - by Bioz Stars, 2026-09
    97/100 stars

    Images

    1) Product Images from "Orthogonal characterization of rAAV reveals vector attributes that drive ITR repair, self-complementary genome formation, and transgene expression"

    Article Title: Orthogonal characterization of rAAV reveals vector attributes that drive ITR repair, self-complementary genome formation, and transgene expression

    Journal: Molecular Therapy. Nucleic Acids

    doi: 10.1016/j.omtn.2026.102899

    Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of extracted rAAV genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with SmaI. (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).
    Figure Legend Snippet: Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of extracted rAAV genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with SmaI. (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).

    Techniques Used: Recombinant, Electrophoresis, Plasmid Preparation, Marker, Expressing, Sequencing, Agarose Gel Electrophoresis

    Related Articles

    Agarose Gel Electrophoresis:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    DNA Extraction:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Transfection:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Plasmid Preparation:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Magnetic Beads:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Protease Inhibitor:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    cDNA Synthesis:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Real-time Polymerase Chain Reaction:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    CCK-8 Assay:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Modification:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Saline:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    In Situ:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Affinity Purification:

    Article Title: hMPV M2-2 protein modulates host mRNA alternative splicing to favor virus replication
    Article Snippet: .. TransStart FastPfu Fly DNA polymerase (TransGen Biotech, Beijing, China; Cat#AP231-22-V4), XhoI (New England Biolabs, Massachusetts, USA; Cat#R0146S), SmaI (New England Biolabs, Massachusetts, USA; CatR0141), rCutSmart buffer (New England Biolabs, Massachusetts, USA; Cat#B6004SVIAL), agarose gel DNA extraction kit (Enhanced) (Tiangen, Beijing, China; Cat#DP219), DH5α competent cells (Weidi Biotechnology, Shanghai, China; Cat#EC1023M), Stbl3 Competent Cells (Weidi Biotechnology, Shanghai, China; Cat#DL1046M), Lipofectamine � RNAiMAX transfection reagent (Thermo Fisher Scientific, Massachusetts, USA; Cat#13778150), plasmid mini kit (Omega Bio-tek, Georgia, USA; Cat#D6904-03), EndoFree midi plasmid kit (Tiangen, Beijing, China; Cat#DP118), Hoechst 33258 (Thermo Fisher Scientific, Massachusetts, USA; Cat#H3569), ANTI-FLAG M2 magnetic beads (SigmaAldrich, Missouri, USA; Cat#M8823-5 mL), 33Flag peptide (GLPBIO, California, USA; Cat#402750-12-3), ProteinSafe� protease inhibitor cocktail (1003) (TransGen Biotech, Beijing, China; Cat#DI111-02), 23 Taq master mix (Vazyme Biotech, Jiangsu, China; Cat#P112-03), TransScript � onestep gDNA removal and cDNA synthesis supermix kit (TransGen Biotech, Beijing, China; Cat#AT311-03), PerfectStart � green qPCR supermix (TransGen Biotech, Beijing, China; Cat#AQ601-04-V2), cell counting kit-8 (Dojindo, Kumamoto Prefecture, Japan; Cat#CK04-500tests), jetOPTIMUS � transfection reagent (Polyplus-transfection, Grand Est, France; Cat#117-07), ampicillin (Sigma-Aldrich, Missouri, USA; Cat#A-9518), Dulbecco’s modified Eagle medium (DMEM) (Gibco, New York, USA; Cat#C11965500BT), Dulbecco’s phosphate -buffered saline (DPBS) (Gibco, New York, USA; Cat#14200075), fetal bovine serum (FBS) (Sigma-Aldrich, Missouri, USA; Cat#F0193), 1% penicillin/ streptomycin antibiotics (Gibco, New York, USA; Cat#15140163), 0.25% trypsin-EDTA (Gibco, New York, USA; Cat#25200114), Duolink � in situ red detection kit (Sigma-Aldrich, Missouri, USA; Cat#DUO92008), affinity-purified donkey anti-rabbit IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92002), affinity-purified donkey anti-mouse IgG (H + L) (Sigma-Aldrich, Missouri, USA; Cat#DUO92004), ProLong gold antifade reagent (Life Technologies, California, USA; Cat#P36930), 4% paraformaldehyde (Biosharp, Anhui, China; Cat#BL539A), BsmBI-v2 (New England Vol. ..

    Control:

    Article Title: Occurrence, antibiotic susceptibility and genetic variation of Streptococcus zooepidemicus in Finnish weanling horses with and without respiratory infection.
    Article Snippet: .. SmaI (New England Biolabs, MA, USA) restriction enzyme patterns of the isolates and control strains were investigated according to a published protocol for Streptococcus pneumoniae [30], with minor modifications: bacterial suspensions of 8.0–9.0 McFarland density were embedded in SeaKem Gold (Lonza, ME, USA) agarose. .. PFGE was performed on a Chef DR III system (Bio-Rad, CA, USA).

    Electrophoresis:

    Article Title: Comprehensive analysis of antimicrobial resistance and virulence determinants in clinical methicillin-resistant Staphylococcus aureus isolates collected in South Korea.
    Article Snippet: The plugs were then washed five times with TE buffer for 20 min each at 55°C using CHEF screened caps (Bio-Rad, Hercules, CA, USA). .. Plug sections were digested with SmaI (New England Biolabs, Ipswich, MA, USA) at 25°C for 4 h. For the reference strain Salmonella enterica ATCC BAA664, plug slices were digested with XbaI (Roche Diagnostics GmbH, Mannheim, Germany) at 37°C for 2.5 h. Digested plug slices were loaded onto 1.2% SeaKem Gold agarose gels (LONZA, Rockland, ME, USA) prepared in 0.5x TBE buffer (Bioneer, Daejeon, Korea) containing 3.75 mM Thiourea (Sigma–Aldrich, St. Louis, MO, USA), and separated using a CHEF Mapper® XA Pulsed Field Electrophoresis System (Bio-Rad, Hercules, CA, USA) under the following conditions: gradient, 6 V/cm; run Time, 18 h; included angle, 120°; initial switch time, 5.16 s; final switch time, 40.17 s; ramping factor, 0. .. Gel images were acquired using the Gel Doc XR+ Gel Documentation system (Bio-Rad, Hercules, CA, USA) and analyzed with BioNumerics version 7.5 (Applied Maths, Sint-Martens-Latem, Belgium).

    Pulsed-Field Gel:

    Article Title: High colonization by multidrug-resistant and virulent Staphylococcus aureus genotypes among critically ill patients in the COVID-19 pandemic in a Brazilian hospital.
    Article Snippet: .. All MRSA isolates were typed by pulsed-field gel electrophoresis (PFGE) after digestion of whole-cell DNA with SmaI and/or ApaI (New England Biolabs; Ipswich, England) in a CHEF-DRIII® system (Bio-Rad, Richmond, CA, USA). ..



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    smai digestion  (New England Biolabs)
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    Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of <t>extracted</t> <t>rAAV</t> genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with <t>SmaI.</t> (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).
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    Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of <t>extracted</t> <t>rAAV</t> genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with <t>SmaI.</t> (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).
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    smai  (New England Biolabs)
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    Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of <t>extracted</t> <t>rAAV</t> genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with <t>SmaI.</t> (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).
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    Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of <t>extracted</t> <t>rAAV</t> genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with <t>SmaI.</t> (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).
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    Image Search Results


    Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of extracted rAAV genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with SmaI. (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Orthogonal characterization of rAAV reveals vector attributes that drive ITR repair, self-complementary genome formation, and transgene expression

    doi: 10.1016/j.omtn.2026.102899

    Figure Lengend Snippet: Recombinant AAV genome element and heterogeneity analysis by native and alkaline electrophoresis (A) Graphic representation of the main recombinant AAV vector genome elements evaluated: wtITR (143 nt, black) versus dITR/wtITR (11-nt deletion in the 5′ C arm, 130 nt, red), EAlbAAT (albumin enhancer-alpha1-antitrypsin promoter, blue), Egfp (enhanced green fluorescent protein, green), pA (bovine/human growth hormone polyadenylation signal, yellow), and CBA (CMV (cytomegalovirus) enhancer-chicken beta actin promoter, orange). Vector genome sizes are shown next to each representation. (B) Native automated electrophoresis of extracted rAAV genomes from dITR/wtITR and wtITR. EalbAAT . Egfp vectors by TapeStation. From left to right: AAV2, AAV3B, AAV5, AAV8, AAV9, and AAVLK03 serotypes. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (C) Native automated electrophoresis of extracted rAAV genomes from AAV5 and AAV8 serotypes with dITR vs. wtITR and CBA versus EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (D) Native automated electrophoresis of extracted rAAV genomes from the AAVLK03 serotype with dITR and CBA vs. EalbAAT promoters. As a run reference, the upper marker (10,000 bp) and lower marker (15 bp) of the ladder are highlighted in purple and green, respectively. (E) Native electrophoresis of extracted rAAV genomes from AAV9 and AAVLK03 serotypes with dITR and CBA versus EalbAAT promoters, before and after digestion at 37°C for 1 h with SmaI. (F) Graphic representation of the dITR/tITR design (11-nt deletion in the 5′ C arm and 22-nt deletion of the 3′ B arm, 121 nt, purple), scITR (C arm deletion, 106 nt, green), and the dITR. AAT . Egfp expression cassette including an hypoxanthine phosphoribosyltransferase 1 ( HPRT 1 ) DNA stuffer sequence (GenBank: NG_012329.2 ) (bottom). Vector genome sizes are shown next to each design. (G) Alkaline electrophoresis of AAV8 vectors with different ITR configurations and CBA versus EalbAAT promoters. (H) Alkaline electrophoresis of AAV8 and AAV3B vectors with dITR/wtITR, dITR/tITR, and scITR configurations. (I) Alkaline electrophoresis of AAV8 and AAVLK03 vectors with the dITR/tITR configuration. (J) Alkaline electrophoresis of AAV3B, AAV5, and AAVLK03 vectors with dITR/wtITR configuration and CBA versus EalbAAT promoters. Combinations are displayed on top of each lane. Agarose gel ladder: FastGene 1 kb DNA marker (10,000–100 bp).

    Article Snippet: SmaI restriction enzyme (20,000 units/mL; New England Biolabs) was used to digest rAAV genomes for 1 h at 37°C.

    Techniques: Recombinant, Electrophoresis, Plasmid Preparation, Marker, Expressing, Sequencing, Agarose Gel Electrophoresis