puc18 Takara Search Results


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  • 85
    TaKaRa puc18 takara vectors
    Evaluation of the specificity of the BLV-CoCoMo-qPCR primers . (A) Real-time PCR using the CoCoMo 6 and CoCoMo 81 primers from the BLV-CoCoMo-qPCR was performed using 0.3 ng of the following infectious molecular clones: BLV (pBLV-IF, lane 2); HTLV-1 (pK30, lane 3); HIV-1 (pNL4-3, lane 4); SIV (pSIVmac239/WT, lane 5); MMTV (hybrid MMTV, lane 6); M-MLV (pL-4, lane 7); and the plasmids <t>pUC18</t> (lane 8), pUC19 (lane 9), pBR322 (lane 10), and pBluescript SK(+) (lane 11). PCR products were subjected to 3% agarose gel electrophoresis. Lane 1, DNA marker Φ × 174- Hae III digest. A PCR product 168 bp in length is indicated by an arrow. (B) The number of BLV provirus copies in 1 μg of DNA from each DNA sample is indicated by lowercase. Values represent the mean ± standard deviation (SD) of the results of three independent experiments.
    Puc18 Takara Vectors, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 48 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa puc18 takara plasmid vector
    WRN deficiency reduces NHEJ. ( a ) Graph indicating in vitro NHEJ efficiency of control and WRN shRNA expression HeLa cell extracts with 2.67 kb cohesive-end ( Sal I-linearized) <t>pUC18</t> DNA substrate. ( b ) NHEJ efficiency of U2OS cell extracts on 5.7 kb non-cohesive DNA substrate derived from Bst XI-digested pSingle-tTS-shRNA plasmid. Graphs represent end-joining efficiency from three independent experiments. *, P value
    Puc18 Takara Plasmid Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc18 takara plasmid vector/product/TaKaRa
    Average 85 stars, based on 20 article reviews
    Price from $9.99 to $1999.99
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    86
    TaKaRa puc18 dna
    Topological inactivation activities. Lane 1 indicated <t>pUC18</t> as a control; Lane 2 indicated the products of pUC18 <t>DNA</t> treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.
    Puc18 Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc18 dna/product/TaKaRa
    Average 86 stars, based on 22 article reviews
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    91
    TaKaRa puc18 based vector harboring egfp
    Topological inactivation activities. Lane 1 indicated <t>pUC18</t> as a control; Lane 2 indicated the products of pUC18 <t>DNA</t> treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.
    Puc18 Based Vector Harboring Egfp, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    TaKaRa puc18 pzy102
    Topological inactivation activities. Lane 1 indicated <t>pUC18</t> as a control; Lane 2 indicated the products of pUC18 <t>DNA</t> treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.
    Puc18 Pzy102, supplied by TaKaRa, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    TaKaRa supercoiled plasmid dna puc18
    Topological inactivation activities. Lane 1 indicated <t>pUC18</t> as a control; Lane 2 indicated the products of pUC18 <t>DNA</t> treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.
    Supercoiled Plasmid Dna Puc18, supplied by TaKaRa, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TaKaRa puc19
    Gel analysis of RNA-primed MPRCA products to detect the contaminating DNA in Phi29 DNA polymerase. ( A ) DNA amplification with Phi29 DNA polymerase prepared by our procedure. ( B ) DNA amplification with commercially available Phi29 DNA polymerase (Epicentre, Lot No. 10710). ( C ) DNA amplification with Phi29 DNA polymerase prepared by DNase-treatment. All gel images show the Bam HI/ Eco RI double digested amplification products. Black arrow indicates the specific amplification products from <t>pUC19;</t> M, AccuRuler 1-kb DNA RTU ladder (Maestrogen). All reactions were repeated three times or more; typical results are shown.
    Puc19, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 476 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    TaKaRa plasmid dna
    Gel analysis of RNA-primed MPRCA products to detect the contaminating DNA in Phi29 DNA polymerase. ( A ) DNA amplification with Phi29 DNA polymerase prepared by our procedure. ( B ) DNA amplification with commercially available Phi29 DNA polymerase (Epicentre, Lot No. 10710). ( C ) DNA amplification with Phi29 DNA polymerase prepared by DNase-treatment. All gel images show the Bam HI/ Eco RI double digested amplification products. Black arrow indicates the specific amplification products from <t>pUC19;</t> M, AccuRuler 1-kb DNA RTU ladder (Maestrogen). All reactions were repeated three times or more; typical results are shown.
    Plasmid Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 92/100, based on 1985 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 1985 article reviews
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    86
    TaKaRa cloning plasmid puc18
    Gel analysis of RNA-primed MPRCA products to detect the contaminating DNA in Phi29 DNA polymerase. ( A ) DNA amplification with Phi29 DNA polymerase prepared by our procedure. ( B ) DNA amplification with commercially available Phi29 DNA polymerase (Epicentre, Lot No. 10710). ( C ) DNA amplification with Phi29 DNA polymerase prepared by DNase-treatment. All gel images show the Bam HI/ Eco RI double digested amplification products. Black arrow indicates the specific amplification products from <t>pUC19;</t> M, AccuRuler 1-kb DNA RTU ladder (Maestrogen). All reactions were repeated three times or more; typical results are shown.
    Cloning Plasmid Puc18, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Evaluation of the specificity of the BLV-CoCoMo-qPCR primers . (A) Real-time PCR using the CoCoMo 6 and CoCoMo 81 primers from the BLV-CoCoMo-qPCR was performed using 0.3 ng of the following infectious molecular clones: BLV (pBLV-IF, lane 2); HTLV-1 (pK30, lane 3); HIV-1 (pNL4-3, lane 4); SIV (pSIVmac239/WT, lane 5); MMTV (hybrid MMTV, lane 6); M-MLV (pL-4, lane 7); and the plasmids pUC18 (lane 8), pUC19 (lane 9), pBR322 (lane 10), and pBluescript SK(+) (lane 11). PCR products were subjected to 3% agarose gel electrophoresis. Lane 1, DNA marker Φ × 174- Hae III digest. A PCR product 168 bp in length is indicated by an arrow. (B) The number of BLV provirus copies in 1 μg of DNA from each DNA sample is indicated by lowercase. Values represent the mean ± standard deviation (SD) of the results of three independent experiments.

    Journal: Retrovirology

    Article Title: BLV-CoCoMo-qPCR: Quantitation of bovine leukemia virus proviral load using the CoCoMo algorithm

    doi: 10.1186/1742-4690-7-91

    Figure Lengend Snippet: Evaluation of the specificity of the BLV-CoCoMo-qPCR primers . (A) Real-time PCR using the CoCoMo 6 and CoCoMo 81 primers from the BLV-CoCoMo-qPCR was performed using 0.3 ng of the following infectious molecular clones: BLV (pBLV-IF, lane 2); HTLV-1 (pK30, lane 3); HIV-1 (pNL4-3, lane 4); SIV (pSIVmac239/WT, lane 5); MMTV (hybrid MMTV, lane 6); M-MLV (pL-4, lane 7); and the plasmids pUC18 (lane 8), pUC19 (lane 9), pBR322 (lane 10), and pBluescript SK(+) (lane 11). PCR products were subjected to 3% agarose gel electrophoresis. Lane 1, DNA marker Φ × 174- Hae III digest. A PCR product 168 bp in length is indicated by an arrow. (B) The number of BLV provirus copies in 1 μg of DNA from each DNA sample is indicated by lowercase. Values represent the mean ± standard deviation (SD) of the results of three independent experiments.

    Article Snippet: Additional clones used included a BLV infectious clone, pBLV-IF [ ]; a HTLV-1 infectious clone, pK30 [ ]; a HIV-1 infectious clone, pNL4-3 [ ]; a SIV infectious clone, SIVmac239/WT [ ]; a hybrid MMTV provirus plasmid [ ]; a M-MLV infectious clone, pL-4 [ ] and plasmids including pUC18 (Takara Bio Inc., Tokyo, Japan), pUC19 (Takara Bio Inc.), and pBR322 (Promega).

    Techniques: Real-time Polymerase Chain Reaction, Clone Assay, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Marker, Standard Deviation

    WRN deficiency reduces NHEJ. ( a ) Graph indicating in vitro NHEJ efficiency of control and WRN shRNA expression HeLa cell extracts with 2.67 kb cohesive-end ( Sal I-linearized) pUC18 DNA substrate. ( b ) NHEJ efficiency of U2OS cell extracts on 5.7 kb non-cohesive DNA substrate derived from Bst XI-digested pSingle-tTS-shRNA plasmid. Graphs represent end-joining efficiency from three independent experiments. *, P value

    Journal: Nature Communications

    Article Title: WRN regulates pathway choice between classical and alternative non-homologous end joining

    doi: 10.1038/ncomms13785

    Figure Lengend Snippet: WRN deficiency reduces NHEJ. ( a ) Graph indicating in vitro NHEJ efficiency of control and WRN shRNA expression HeLa cell extracts with 2.67 kb cohesive-end ( Sal I-linearized) pUC18 DNA substrate. ( b ) NHEJ efficiency of U2OS cell extracts on 5.7 kb non-cohesive DNA substrate derived from Bst XI-digested pSingle-tTS-shRNA plasmid. Graphs represent end-joining efficiency from three independent experiments. *, P value

    Article Snippet: In vitro NHEJ assays were performed as before with DNA substrates derived from Sal I linearized pUC18 plasmid (cohesive ends) and from 5.7 kb non-cohesive substrate from I-Sce I digested pSingle-tTS-plasmid (Clontech).

    Techniques: Non-Homologous End Joining, In Vitro, shRNA, Expressing, Derivative Assay, Plasmid Preparation

    Schematic diagrams of the plasmids. (A) pUC18-HVT-TK. The region from 45,700 to 48,967 nucleotides (nts) of the herpesvirus of turkeys (HVT) FC126 strain genome was cloned into pUC18. (B) pUC18-HVT-TK- Sfi I. The Sfi I recognition site was introduced between nts 47,316 and 47,317 of the FC126 genome, and the 45,700–48,967 region was cloned into pUC18. (C) pUC18-HVT-TK-I- Sce I- Sfi I. A 50-bp duplication sequence (nts 47,317–47,366 of the FC126 genome) and the I- Sce I recognition site were inserted before the Sfi I site. Dashed lines show homologous sequences. (D) pUC18-HVT-BAC. LoxP , eGFP, mini-F and chloramphenicol resistance cassette sequences were inserted into the Sfi I site of pUC18-HVT-TK-I- Sce I- Sfi I. Dashed lines show homologous sequences. Cm R indicates the chloramphenicol resistance gene.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Removal of Inserted BAC after linearizatiON (RIBON)—a novel strategy to excise the mini-F sequences from viral BAC vectors

    doi: 10.1292/jvms.16-0038

    Figure Lengend Snippet: Schematic diagrams of the plasmids. (A) pUC18-HVT-TK. The region from 45,700 to 48,967 nucleotides (nts) of the herpesvirus of turkeys (HVT) FC126 strain genome was cloned into pUC18. (B) pUC18-HVT-TK- Sfi I. The Sfi I recognition site was introduced between nts 47,316 and 47,317 of the FC126 genome, and the 45,700–48,967 region was cloned into pUC18. (C) pUC18-HVT-TK-I- Sce I- Sfi I. A 50-bp duplication sequence (nts 47,317–47,366 of the FC126 genome) and the I- Sce I recognition site were inserted before the Sfi I site. Dashed lines show homologous sequences. (D) pUC18-HVT-BAC. LoxP , eGFP, mini-F and chloramphenicol resistance cassette sequences were inserted into the Sfi I site of pUC18-HVT-TK-I- Sce I- Sfi I. Dashed lines show homologous sequences. Cm R indicates the chloramphenicol resistance gene.

    Article Snippet: The resultant 3.3-kbp fragment was cloned into the pUC18 vector (Takara) digested with Sal I and Sac I, resulting in pUC18-HVT-TK-Sfi I ( ).

    Techniques: Clone Assay, Sequencing, BAC Assay

    Plaques produced by reconstituted HVT- Sfi I in chicken embryo fibroblasts. Cells were transfected with linearized pHVT-BAC and pUC18-HVT-TK- Sfi I and analyzed for plaque formation and eGFP expression five days after transfection. Left panels, eGFP fluorescence; right panels, bright field microscopy. Scale bars, 50 µ m.

    Journal: The Journal of Veterinary Medical Science

    Article Title: Removal of Inserted BAC after linearizatiON (RIBON)—a novel strategy to excise the mini-F sequences from viral BAC vectors

    doi: 10.1292/jvms.16-0038

    Figure Lengend Snippet: Plaques produced by reconstituted HVT- Sfi I in chicken embryo fibroblasts. Cells were transfected with linearized pHVT-BAC and pUC18-HVT-TK- Sfi I and analyzed for plaque formation and eGFP expression five days after transfection. Left panels, eGFP fluorescence; right panels, bright field microscopy. Scale bars, 50 µ m.

    Article Snippet: The resultant 3.3-kbp fragment was cloned into the pUC18 vector (Takara) digested with Sal I and Sac I, resulting in pUC18-HVT-TK-Sfi I ( ).

    Techniques: Produced, Transfection, BAC Assay, Expressing, Fluorescence, Microscopy

    The effect of S1 nuclease on the PCR product. The PCR product, as well as M13mp18 (single-stranded) DNA and Sma I digests of pUC18 DNA, were digested with S1 nuclease. The top numbers indicate the amounts of S1 nuclease added per tube. DNAs were separated by 0.8% agarose gel electrophoresis under neutral conditions as in the legend to Figure 1.

    Journal: Nucleic Acids Research

    Article Title: Telomere repeat DNA forms a large non-covalent complex with unique cohesive properties which is dissociated by Werner syndrome DNA helicase in the presence of replication protein A

    doi:

    Figure Lengend Snippet: The effect of S1 nuclease on the PCR product. The PCR product, as well as M13mp18 (single-stranded) DNA and Sma I digests of pUC18 DNA, were digested with S1 nuclease. The top numbers indicate the amounts of S1 nuclease added per tube. DNAs were separated by 0.8% agarose gel electrophoresis under neutral conditions as in the legend to Figure 1.

    Article Snippet: The PCR product, as well as Sma I digests of plasmid pUC18 (Takara) as a reference for double-stranded DNA and plasmid M13mp18 (Takara) as a reference for single-stranded DNA, were used as DNA substrates.

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis

    Topological inactivation activities. Lane 1 indicated pUC18 as a control; Lane 2 indicated the products of pUC18 DNA treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.

    Journal: PLoS ONE

    Article Title: A Novel Method for Simultaneous Production of Two Ribosome-Inactivating Proteins, ?-MMC and MAP30, from Momordica charantia L

    doi: 10.1371/journal.pone.0101998

    Figure Lengend Snippet: Topological inactivation activities. Lane 1 indicated pUC18 as a control; Lane 2 indicated the products of pUC18 DNA treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.

    Article Snippet: LMW Calibration Kit was supplied by SIBAS (Shanghai, China). pUC18 DNA used in detection of topological activity was obtained from TAKARA (Dalian, China).

    Techniques:

    Representative ultraviolet images from ethidium bromide-stained PAGE gels of the 110 nt PCR product derived from pUC18 (amplifying region I) and the C5-modified nucleoside triphosphates. ( A–C ) The reaction mixtures containing dATP, dGTP, dCTP and T AL (lane 3), T AC (lane 4), T AF (lane 5), T PA (lane 6), T PN (lane 7), T PR (lane 8), T A2 (lane 11), T A4 (lane 12), T A6 (lane 13), T G6 (lane 14), T ME (lane 15), T CN (lane 16), or T DH (lane 17) were separated by electrophoresis on denaturing PAGE. Except for the positive control, the reaction mixtures do not contain natural TTP. The PCR product was generated by the positive control (reaction mixture containing dATP, dGTP, dCTP and TTP; lanes 2 and 10) but not by the negative control (reaction mixture containing dATP, dGTP and dCTP; lanes 1 and 9). ( D–F ) Reaction mixtures containing dATP, dGTP, TTP and C AL (lane 3), C AC (lane 4), C AF (lane 5), C PA (lane 6), C PN (lane 7), C PR (lane 8), C A2 (lane 11), C A4 (lane 12), C A6 (lane 13), C G6 (lane 14), C ME (lane 15), C CN (lane 16) or C DH (lane 17) were separated by electrophoresis on denaturing PAGE. Except for the positive control, the reaction mixtures did not contain natural dCTP. The PCR product was generated by the positive control (reaction mixture containing dATP, dGTP, dCTP and TTP; lanes 2 and 10) but not by the negative control (reaction mixture containing dATP, dGTP and TTP; lanes 1 and 9). The thermostable DNA polymerases used were Taq (A, D), Vent(exo-) (B and E), and KOD Dash (C and F).

    Journal: Nucleic Acids Research

    Article Title: Systematic characterization of 2?-deoxynucleoside- 5?-triphosphate analogs as substrates for DNA polymerases by polymerase chain reaction and kinetic studies on enzymatic production of modified DNA

    doi: 10.1093/nar/gkl637

    Figure Lengend Snippet: Representative ultraviolet images from ethidium bromide-stained PAGE gels of the 110 nt PCR product derived from pUC18 (amplifying region I) and the C5-modified nucleoside triphosphates. ( A–C ) The reaction mixtures containing dATP, dGTP, dCTP and T AL (lane 3), T AC (lane 4), T AF (lane 5), T PA (lane 6), T PN (lane 7), T PR (lane 8), T A2 (lane 11), T A4 (lane 12), T A6 (lane 13), T G6 (lane 14), T ME (lane 15), T CN (lane 16), or T DH (lane 17) were separated by electrophoresis on denaturing PAGE. Except for the positive control, the reaction mixtures do not contain natural TTP. The PCR product was generated by the positive control (reaction mixture containing dATP, dGTP, dCTP and TTP; lanes 2 and 10) but not by the negative control (reaction mixture containing dATP, dGTP and dCTP; lanes 1 and 9). ( D–F ) Reaction mixtures containing dATP, dGTP, TTP and C AL (lane 3), C AC (lane 4), C AF (lane 5), C PA (lane 6), C PN (lane 7), C PR (lane 8), C A2 (lane 11), C A4 (lane 12), C A6 (lane 13), C G6 (lane 14), C ME (lane 15), C CN (lane 16) or C DH (lane 17) were separated by electrophoresis on denaturing PAGE. Except for the positive control, the reaction mixtures did not contain natural dCTP. The PCR product was generated by the positive control (reaction mixture containing dATP, dGTP, dCTP and TTP; lanes 2 and 10) but not by the negative control (reaction mixture containing dATP, dGTP and TTP; lanes 1 and 9). The thermostable DNA polymerases used were Taq (A, D), Vent(exo-) (B and E), and KOD Dash (C and F).

    Article Snippet: Primers 1F, 1R, 2F and 2R were purchased from JBioS, and pUC18 template DNA was obtained from TaKaRa Biomedicals.

    Techniques: Staining, Polyacrylamide Gel Electrophoresis, Polymerase Chain Reaction, Derivative Assay, Modification, Electrophoresis, Positive Control, Generated, Negative Control

    Sequences of amplifying regions of the pUC18 template DNA. Primer sequences are underlined.

    Journal: Nucleic Acids Research

    Article Title: Systematic characterization of 2?-deoxynucleoside- 5?-triphosphate analogs as substrates for DNA polymerases by polymerase chain reaction and kinetic studies on enzymatic production of modified DNA

    doi: 10.1093/nar/gkl637

    Figure Lengend Snippet: Sequences of amplifying regions of the pUC18 template DNA. Primer sequences are underlined.

    Article Snippet: Primers 1F, 1R, 2F and 2R were purchased from JBioS, and pUC18 template DNA was obtained from TaKaRa Biomedicals.

    Techniques:

    Gel analysis of RNA-primed MPRCA products to detect the contaminating DNA in Phi29 DNA polymerase. ( A ) DNA amplification with Phi29 DNA polymerase prepared by our procedure. ( B ) DNA amplification with commercially available Phi29 DNA polymerase (Epicentre, Lot No. 10710). ( C ) DNA amplification with Phi29 DNA polymerase prepared by DNase-treatment. All gel images show the Bam HI/ Eco RI double digested amplification products. Black arrow indicates the specific amplification products from pUC19; M, AccuRuler 1-kb DNA RTU ladder (Maestrogen). All reactions were repeated three times or more; typical results are shown.

    Journal: PLoS ONE

    Article Title: Preparation of Phi29 DNA Polymerase Free of Amplifiable DNA Using Ethidium Monoazide, an Ultraviolet-Free Light-Emitting Diode Lamp and Trehalose

    doi: 10.1371/journal.pone.0082624

    Figure Lengend Snippet: Gel analysis of RNA-primed MPRCA products to detect the contaminating DNA in Phi29 DNA polymerase. ( A ) DNA amplification with Phi29 DNA polymerase prepared by our procedure. ( B ) DNA amplification with commercially available Phi29 DNA polymerase (Epicentre, Lot No. 10710). ( C ) DNA amplification with Phi29 DNA polymerase prepared by DNase-treatment. All gel images show the Bam HI/ Eco RI double digested amplification products. Black arrow indicates the specific amplification products from pUC19; M, AccuRuler 1-kb DNA RTU ladder (Maestrogen). All reactions were repeated three times or more; typical results are shown.

    Article Snippet: As a result, no amplification product were found in plural NTCs using the purified Phi29 DNA polymerase, while 2.7 kb fragment was found in all positive controls using pUC19 as a template DNA ( ).

    Techniques: Amplification

    Topoisomerase I relaxes negative supercoiled plasmid DNA in present RstA. Lane 1, 0.4 μg pUC19 plasmid with no added protein. Lane 2, 0.4 μg pUC19 plasmid with BSA protein. And the other lanes contain the same concentrations of plasmid as lane 2, but different amounts of purified protein. Lanes 3–8 contain decreasing amounts of purified RstA protein (from 500 ng to 0 μg) and one unit Topoisomerase I. Lane 9, 500 ng purified RstA protein without Topoisomerase I. 6×loading buffer was added to stop the reaction and analyzed by 0.8% agrose gel electrophoresis.

    Journal: PLoS ONE

    Article Title: Absence of RstA results in delayed initiation of DNA replication in Escherichia coli

    doi: 10.1371/journal.pone.0200688

    Figure Lengend Snippet: Topoisomerase I relaxes negative supercoiled plasmid DNA in present RstA. Lane 1, 0.4 μg pUC19 plasmid with no added protein. Lane 2, 0.4 μg pUC19 plasmid with BSA protein. And the other lanes contain the same concentrations of plasmid as lane 2, but different amounts of purified protein. Lanes 3–8 contain decreasing amounts of purified RstA protein (from 500 ng to 0 μg) and one unit Topoisomerase I. Lane 9, 500 ng purified RstA protein without Topoisomerase I. 6×loading buffer was added to stop the reaction and analyzed by 0.8% agrose gel electrophoresis.

    Article Snippet: Assay of topoisomerase I activity Add 2 μl of 10×topoisomerase I reaction buffer and 400 ng pUC19 plasmid DNA (Takara, Japan) to each of a series of 1.5-ml microcentrifuge tubes on ice.

    Techniques: Plasmid Preparation, Purification, Nucleic Acid Electrophoresis