bglii Takara Search Results


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  • 99
    New England Biolabs bgl ii
    Bgl Ii, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1668 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher clonejet pcr cloning kit
    Clonejet Pcr Cloning Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8129 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa bglii
    Schematic diagram of vector construction. 1, PCR products of upstream homologous arm 2A with AscI and <t>BglII,</t> cloned into the <t>pMD18-T</t> vector and named pMD18-T-2A. pMD18-T-2A was digested with AscI and BglII endonucleases and inserted into the pGBHCupp
    Bglii, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 2129 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa ecori
    Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, <t>EcoRI</t> and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with <t>BamHI,</t> BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.
    Ecori, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 8219 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa bamhi
    Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, <t>EcoRI</t> and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with <t>BamHI,</t> BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.
    Bamhi, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 6140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nhei  (TaKaRa)
    99
    TaKaRa nhei
    Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, <t>EcoRI</t> and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with <t>BamHI,</t> BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.
    Nhei, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1827 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    xhoi  (TaKaRa)
    99
    TaKaRa xhoi
    Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, <t>EcoRI</t> and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with <t>BamHI,</t> BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.
    Xhoi, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 5928 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa hindiii
    Identification of the recombinant plasmid POT1-promoter-2. (A) Electrophoresis of BglII and <t>HindIII</t> digested products. M, molecular weight marker (200–5,000 bp); lane 1 and 2, enzyme digestion products (5,057 and 461 bp). (B) Electrophoresis of the PCR products. M, molecular weight marker (100–600 bp); lane 1–4, the PCR products (461 bp) of the POT1-promoter-2 plasmid using primers for POT1-promoter-2. (C) Sequencing results of POT1-promoter-2. POT1, protection of telomere 1; PCR, polymerase chain reaction; bp, base pairs.
    Hindiii, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 3811 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pcr amplification
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    Pcr Amplification, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 21524 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa in fusion hd cloning kit
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    In Fusion Hd Cloning Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 15962 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    TaKaRa t4 dna ligase
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    T4 Dna Ligase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 7380 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    kpni  (TaKaRa)
    99
    TaKaRa kpni
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    Kpni, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1808 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pmcherry c1
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    Pmcherry C1, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1577 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs t4 dna ligase
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    T4 Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 49148 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa pcmv myc vector
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    Pcmv Myc Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1196 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa plvx tight puro
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    Plvx Tight Puro, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 528 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa la taq dna polymerase
    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative <t>PCR</t> for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing <t>TFEB-GFP</t> with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P
    La Taq Dna Polymerase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 5238 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    xbai  (TaKaRa)
    99
    TaKaRa xbai
    CRISPR-Cas9 enhances KI efficiency in cjESCs. ( a ) Schematic diagram of the ACTB-EGFP system. The ACTB-EGFP TV harboured IRES-EGFP-2A-Neo flanked by 2.5-kb and 5.5-kb homology arms to the surrounding regions of the 3′-UTR of the marmoset ACTB gene locus. The three gRNAs (ACTB-1, 2, 3, their recognition sites are shown as scissors) target the 3′-UTR region, which is not included in the TV. The TV is not detected by the gRNAs for the marmoset ACTB gene. Black thin arrows show the primer binding sites for genotyping PCR; x, a restriction enzyme site ( <t>XbaI</t> ); p, 5′-external probe for Southern blotting. ( b ) The number of G418-resistant colonies following G418 selection of 1 × 10 6 transfected ESCs, shown as the mean + s.e.m., n = 3. The number of colonies with strong EGFP fluorescence (EGFP++) is shown in dark grey; the number of colonies with moderate EGFP fluorescence (EGFP+) is shown in grey; the number of EGFP-negative colonies (EGFP−) is shown in bright grey. ( c ) A representative image of an EGFP++ (left) and EGFP+ (right) colony observed under bright field (BF) or under green fluorescence after G418 selection. Scale bar, 200 μm. ( d ) Genotyping PCR analysis of EGFP++ and EGFP+ cjESC clones. M, <t>DNA</t> marker. The separate images were cropped from the same gel. ( e ) Southern blotting analysis of EGFP++ and EGFP+ clones using the 5′-external probe. M, DNA marker. The separate images were cropped from the same gel. The entire image of the gel is shown in Supplementary Fig. S14a . ( f ) Schematic diagram of the shortened ACTB-EGFP TVs. ( g ) The number of G418-resistant colonies following selection of 1 × 10 6 transfected cjESCs, shown as the mean + s.e.m., n = 3. Each group is represented by the same colours as in ( b ). * P
    Xbai, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1293 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Schematic diagram of vector construction. 1, PCR products of upstream homologous arm 2A with AscI and BglII, cloned into the pMD18-T vector and named pMD18-T-2A. pMD18-T-2A was digested with AscI and BglII endonucleases and inserted into the pGBHCupp

    Journal: Applied and Environmental Microbiology

    Article Title: Chromosomal Insertions in the Lactobacillus caseiupp Gene That Are Useful for Vaccine Expression

    doi: 10.1128/AEM.00175-14

    Figure Lengend Snippet: Schematic diagram of vector construction. 1, PCR products of upstream homologous arm 2A with AscI and BglII, cloned into the pMD18-T vector and named pMD18-T-2A. pMD18-T-2A was digested with AscI and BglII endonucleases and inserted into the pGBHCupp

    Article Snippet: The pMD18-T-2A plasmid was digested with BglII (TaKaRa) and AscI (NEB).

    Techniques: Plasmid Preparation, Polymerase Chain Reaction, Clone Assay

    Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.

    Journal: Emerging Microbes & Infections

    Article Title: Comparative genomic analysis of two emergent human adenovirus type 14 respiratory pathogen isolates in China reveals similar yet divergent genomes

    doi: 10.1038/emi.2017.78

    Figure Lengend Snippet: Identification of the genome types of HAdV-B14 strains GZ01 (Guangzhou), BJ430 (Beijing), and CHN2012 (Beijing). The in silico restriction enzyme analysis (REA) digestion patterns of HAdV-14 genomes were generated using the Vector NTI 10.3.0 software (Invitrogen Corp., San Diego, CA, USA). Genome digestion patterns of the strains are as follows: ( A ) de Wit or prototype (AY803294); ( B ) GZ01 (JQ824845); ( C ) 303600, reported as HAdV-B14p1 (FJ822614); ( D ) BJ430 (JN032132); and ( E ) CHN2012 (JX892927). These were analyzed with Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal I, Sma I, Xba I, and Xho I, as described by Li et al. 59 All the three recent China outbreak genomes correspond to the genome type of HAdV-14p1 and have identical REA profiles to each other. They are divergent from the HAdV-B14 prototype (1955). M: 1 kb DNA Sizing Ladder.

    Article Snippet: Genome type identification In silico restriction maps of available genome sequences from all HAdV-14 strains CHN/GZ01/2010, 303600, CHN/BJ430, CHN2012, and prototype de Wit were generated using the Vector NTI 10.3.0 software (Invitrogen Corp.; San Diego, CA, USA) as described in earlier studies., , These included profiles from restriction enzymes Bam HI, Bcl I, Bgl l, Bgl II, Bst EII, Eco RI, Hin dIII, Hpa I, Sal l, Sma I, Xba I, and Xho I (TaKaRa Corp.; China), all of which were chosen in order to be consistent with the original nomenclature system devised by Li and colleagues.

    Techniques: In Silico, Generated, Plasmid Preparation, Software

    In vitro MMR efficiency of BS cell lines. ( A ) Western blot showing the absence of BLM protein in the BS cell lines. Aliquots of 25 µg of the indicated nuclear extracts were probed with anti-BLM antibody (IHIC33) and anti-XPB antibody [TFIIH p89 (S-19); Santa Cruz Biotechnology] as control. ( B ) MMR efficiency of the BLM-negative human fibroblast GM08505 and of the human lymphoblasts GM09960 and GM03403. The MMR-proficient MRC5 SV40 fibroblasts, TK6 lymphoblasts, PSNF5 cells (GM08505 stably transfected with BLM cDNA) and the MMR-deficient hMLH1 –/– colon cancer cell line HCT116 were used as controls. The MMR efficiencies of the two BS lymphoblasts complemented with purified recombinant BLM protein are also shown. The repair efficiency is expressed as fmol phagemid DNA cleaved by Bgl II in 30 min.

    Journal: Nucleic Acids Research

    Article Title: Direct association of Bloom's syndrome gene product with the human mismatch repair protein MLH1

    doi:

    Figure Lengend Snippet: In vitro MMR efficiency of BS cell lines. ( A ) Western blot showing the absence of BLM protein in the BS cell lines. Aliquots of 25 µg of the indicated nuclear extracts were probed with anti-BLM antibody (IHIC33) and anti-XPB antibody [TFIIH p89 (S-19); Santa Cruz Biotechnology] as control. ( B ) MMR efficiency of the BLM-negative human fibroblast GM08505 and of the human lymphoblasts GM09960 and GM03403. The MMR-proficient MRC5 SV40 fibroblasts, TK6 lymphoblasts, PSNF5 cells (GM08505 stably transfected with BLM cDNA) and the MMR-deficient hMLH1 –/– colon cancer cell line HCT116 were used as controls. The MMR efficiencies of the two BS lymphoblasts complemented with purified recombinant BLM protein are also shown. The repair efficiency is expressed as fmol phagemid DNA cleaved by Bgl II in 30 min.

    Article Snippet: The yeast strain L40 [ MATa trp1 leu2 his3 LYS2 :: lexA-HIS3 URA3 :: lexA-lacZ ] was sequentially transformed with the bait pBTM116-BLM (amino acids 770–1417) and a random primed human peripheral blood cDNA library cloned into the Bgl II sites of pACT (Clonetech) using the lithium acetate method.

    Techniques: In Vitro, Western Blot, Stable Transfection, Transfection, Purification, Recombinant

    Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, EcoRI and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with BamHI, BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.

    Journal: PLoS ONE

    Article Title: Lacanobia oleracea nucleopolyhedrovirus (LaolNPV): A new European species of alphabaculovirus with a narrow host range

    doi: 10.1371/journal.pone.0176171

    Figure Lengend Snippet: Restriction endonuclease analysis of LaolNPV DNA. Restriction endonuclease profiles following digestion of A) genomic DNAs of the seven isolates of LaolNPV collected in alfalfa crops nearby Montpellier with BglII, EcoRI and PstI enzymes, B) genomic DNAs of Chrysodeixis chalcites NPV (Chch), Helicoverpa armigera SNPV (Hear), Spodoptera littoralis (Spli), Spodopera exigua MNPV (Se) and the virus under study (LaolNPV) with BamHI, BglII, EcoRI and PstI, and C) genomic DNAs of Mamestra brassicae MNPV (Mb), Mamestra configurata NPV-A (MacoA), Mamestra configurata NPV-B (MacoB) and LaolNPV with BglII, EcoRI and PstI.

    Article Snippet: For restriction endonuclease (REN) digestion, a sample of viral DNA (1–2 μg) was incubated with one of the following enzymes BamHI, BglII, EcoRI or PstI (Takara) (10 U) at 37°C for 4–12 h. Each reaction was stopped by the addition of 4 μl of 6x loading buffer (0.25% w/v bromophenol blue and 40% w/v sucrose).

    Techniques:

    Identification of the recombinant plasmid POT1-promoter-2. (A) Electrophoresis of BglII and HindIII digested products. M, molecular weight marker (200–5,000 bp); lane 1 and 2, enzyme digestion products (5,057 and 461 bp). (B) Electrophoresis of the PCR products. M, molecular weight marker (100–600 bp); lane 1–4, the PCR products (461 bp) of the POT1-promoter-2 plasmid using primers for POT1-promoter-2. (C) Sequencing results of POT1-promoter-2. POT1, protection of telomere 1; PCR, polymerase chain reaction; bp, base pairs.

    Journal: Oncology Letters

    Article Title: Construction of the POT1 promoter report gene vector, and the effect and underlying mechanism of the POT1 promoter in regulating telomerase and telomere length

    doi: 10.3892/ol.2017.7127

    Figure Lengend Snippet: Identification of the recombinant plasmid POT1-promoter-2. (A) Electrophoresis of BglII and HindIII digested products. M, molecular weight marker (200–5,000 bp); lane 1 and 2, enzyme digestion products (5,057 and 461 bp). (B) Electrophoresis of the PCR products. M, molecular weight marker (100–600 bp); lane 1–4, the PCR products (461 bp) of the POT1-promoter-2 plasmid using primers for POT1-promoter-2. (C) Sequencing results of POT1-promoter-2. POT1, protection of telomere 1; PCR, polymerase chain reaction; bp, base pairs.

    Article Snippet: Subsequently, the pGL3-control plasmid (Promega Corporation, Madison, WI, USA) and PCR products containing the desired sequence were double-digested with BglII and HindIII , and annealed together using T4 DNA ligase (Takara Biotechnology Co., Ltd.) at 16°C overnight.

    Techniques: Recombinant, Plasmid Preparation, Electrophoresis, Molecular Weight, Marker, Polymerase Chain Reaction, Sequencing

    Identification of the recombinant plasmid POT1-promoter-1. (A) Electrophoresis of BglII and HindIII digested products. M, molecular weight marker (200–5,000 bp); lane 1–3, enzyme digestion products (5,057 and 207 bp). (B) Electrophoresis of the PCR products. M, molecular weight marker (100–600 bp); lane 1–4, the PCR products (207 bp) of the POT1-promoter-1 plasmid using primers for POT1-promoter-1. (C) Sequencing results of POT1-promoter-1. POT1, protection of telomere 1; PCR, polymerase chain reaction; bp, base pairs.

    Journal: Oncology Letters

    Article Title: Construction of the POT1 promoter report gene vector, and the effect and underlying mechanism of the POT1 promoter in regulating telomerase and telomere length

    doi: 10.3892/ol.2017.7127

    Figure Lengend Snippet: Identification of the recombinant plasmid POT1-promoter-1. (A) Electrophoresis of BglII and HindIII digested products. M, molecular weight marker (200–5,000 bp); lane 1–3, enzyme digestion products (5,057 and 207 bp). (B) Electrophoresis of the PCR products. M, molecular weight marker (100–600 bp); lane 1–4, the PCR products (207 bp) of the POT1-promoter-1 plasmid using primers for POT1-promoter-1. (C) Sequencing results of POT1-promoter-1. POT1, protection of telomere 1; PCR, polymerase chain reaction; bp, base pairs.

    Article Snippet: Subsequently, the pGL3-control plasmid (Promega Corporation, Madison, WI, USA) and PCR products containing the desired sequence were double-digested with BglII and HindIII , and annealed together using T4 DNA ligase (Takara Biotechnology Co., Ltd.) at 16°C overnight.

    Techniques: Recombinant, Plasmid Preparation, Electrophoresis, Molecular Weight, Marker, Polymerase Chain Reaction, Sequencing

    TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative PCR for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing TFEB-GFP with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P

    Journal: The Journal of Cell Biology

    Article Title: MiT/TFE transcription factors are activated during mitophagy downstream of Parkin and Atg5

    doi: 10.1083/jcb.201501002

    Figure Lengend Snippet: TFEB translocates to the nucleus during mitophagy in a Parkin- and PINK1-dependent manner. (A) YFP-Parkin HeLa cells were treated with O/A for up to 10 h, fractionated, and immunoblotted. (B) Quantification of data in A. Endogenous TFEB expression was normalized to GAPDH (cytosol) or histone H3 (nuclear) and nuclear TFEB expressed as a percentage of total TFEB. Data are means ± SD ( n = 3). (C) mCherry-Parkin HeLa cells were left untreated (Control), starved (2 h), or treated with torin 1 (2 h), O/A (6 h), or valinomycin (Val; 6 h). CIP treatment of cell lysates was performed before immunoblotting. (D) WT and mCherry-Parkin HeLa cells were treated with DMSO or O/A (6 h), lysed, and immunoblotted. A CIP-treated control was included as a reference for total TFEB dephosphorylation. (E) WT and mCherry-Parkin HeLa cells were treated with DMSO, torin 1, or O/A for 18 h and analyzed by quantitative PCR for TFEB target gene expression. Data are means ± SD ( n = 3). (F) WT and PINK1 KO HeLa cells stably expressing TFEB-GFP with or without mCherry-Parkin were treated as in C. Fixed cells were stained with DAPI and analyzed by immunofluorescence. Bars, 10 µm. See Fig. S1 F for quantification. *, P

    Article Snippet: The viral TFEC-GFP expression vector was generated in a two-step process through PCR amplification of the full-length TFEC encoding sequence from pCMV6-Entry-TFEC-Myc-DDK followed by in-frame cloning into XhoI–BamHI sites of the pEYFP-N1 vector (Takara Bio Inc.).

    Techniques: Expressing, De-Phosphorylation Assay, Real-time Polymerase Chain Reaction, Stable Transfection, Staining, Immunofluorescence

    CRISPR-Cas9 enhances KI efficiency in cjESCs. ( a ) Schematic diagram of the ACTB-EGFP system. The ACTB-EGFP TV harboured IRES-EGFP-2A-Neo flanked by 2.5-kb and 5.5-kb homology arms to the surrounding regions of the 3′-UTR of the marmoset ACTB gene locus. The three gRNAs (ACTB-1, 2, 3, their recognition sites are shown as scissors) target the 3′-UTR region, which is not included in the TV. The TV is not detected by the gRNAs for the marmoset ACTB gene. Black thin arrows show the primer binding sites for genotyping PCR; x, a restriction enzyme site ( XbaI ); p, 5′-external probe for Southern blotting. ( b ) The number of G418-resistant colonies following G418 selection of 1 × 10 6 transfected ESCs, shown as the mean + s.e.m., n = 3. The number of colonies with strong EGFP fluorescence (EGFP++) is shown in dark grey; the number of colonies with moderate EGFP fluorescence (EGFP+) is shown in grey; the number of EGFP-negative colonies (EGFP−) is shown in bright grey. ( c ) A representative image of an EGFP++ (left) and EGFP+ (right) colony observed under bright field (BF) or under green fluorescence after G418 selection. Scale bar, 200 μm. ( d ) Genotyping PCR analysis of EGFP++ and EGFP+ cjESC clones. M, DNA marker. The separate images were cropped from the same gel. ( e ) Southern blotting analysis of EGFP++ and EGFP+ clones using the 5′-external probe. M, DNA marker. The separate images were cropped from the same gel. The entire image of the gel is shown in Supplementary Fig. S14a . ( f ) Schematic diagram of the shortened ACTB-EGFP TVs. ( g ) The number of G418-resistant colonies following selection of 1 × 10 6 transfected cjESCs, shown as the mean + s.e.m., n = 3. Each group is represented by the same colours as in ( b ). * P

    Journal: Scientific Reports

    Article Title: Robust and efficient knock-in in embryonic stem cells and early-stage embryos of the common marmoset using the CRISPR-Cas9 system

    doi: 10.1038/s41598-018-37990-w

    Figure Lengend Snippet: CRISPR-Cas9 enhances KI efficiency in cjESCs. ( a ) Schematic diagram of the ACTB-EGFP system. The ACTB-EGFP TV harboured IRES-EGFP-2A-Neo flanked by 2.5-kb and 5.5-kb homology arms to the surrounding regions of the 3′-UTR of the marmoset ACTB gene locus. The three gRNAs (ACTB-1, 2, 3, their recognition sites are shown as scissors) target the 3′-UTR region, which is not included in the TV. The TV is not detected by the gRNAs for the marmoset ACTB gene. Black thin arrows show the primer binding sites for genotyping PCR; x, a restriction enzyme site ( XbaI ); p, 5′-external probe for Southern blotting. ( b ) The number of G418-resistant colonies following G418 selection of 1 × 10 6 transfected ESCs, shown as the mean + s.e.m., n = 3. The number of colonies with strong EGFP fluorescence (EGFP++) is shown in dark grey; the number of colonies with moderate EGFP fluorescence (EGFP+) is shown in grey; the number of EGFP-negative colonies (EGFP−) is shown in bright grey. ( c ) A representative image of an EGFP++ (left) and EGFP+ (right) colony observed under bright field (BF) or under green fluorescence after G418 selection. Scale bar, 200 μm. ( d ) Genotyping PCR analysis of EGFP++ and EGFP+ cjESC clones. M, DNA marker. The separate images were cropped from the same gel. ( e ) Southern blotting analysis of EGFP++ and EGFP+ clones using the 5′-external probe. M, DNA marker. The separate images were cropped from the same gel. The entire image of the gel is shown in Supplementary Fig. S14a . ( f ) Schematic diagram of the shortened ACTB-EGFP TVs. ( g ) The number of G418-resistant colonies following selection of 1 × 10 6 transfected cjESCs, shown as the mean + s.e.m., n = 3. Each group is represented by the same colours as in ( b ). * P

    Article Snippet: For digestion of genomic DNA, we used XbaI (ACTB-EGFP ), BglII (PLP1-EGFP ) and EcoRV (FOXP2 , PLP1 -P216S, S253T and A39T) (purchased from Takara or NEB).

    Techniques: CRISPR, Binding Assay, Polymerase Chain Reaction, Southern Blot, Selection, Transfection, Fluorescence, Clone Assay, Marker