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  • 79
    Thermo Fisher ptz 57r t vector
    Map of PTZ 57 R/T-VP7 clone (designed with snap gene software): Sequence analysis indicates that the PTZ57 R/T-VP7 Clone had a full-length copy of segment 9 (VP7) of human rotavirus, and showed 99% homology with RV4 strain (GenBank Sequence ID: M64666.1). The  Bstu-1  restriction enzyme cleaves the PTZ57 R/T-VP7 Clone in the multiple positions. Two of these positions resulted in 635 and 1733 fragments that ultimately released the full-length of VP7 sequence from the plasmid
    Ptz 57r T Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Thermo Fisher ptz57r
    Graphic Comparison of Colony Counting and OD 600 Measurement Results Between Case and Control Groups The dot plot graph of colony counting results; A, and OD600 measurement results; B from the three transformation procedures performed at two different incubation temperatures: 30 and 37°C. The data shown in the graphs reveal a significant reduction in colony numbers and bacterial growth in the E. coli TOP10 containing pZFN, transformed with <t>pTZ57R,</t> compared to the E. coli TOP10 containing the pP15A, kana R vector transformed with pTZ57R, on amp and amp-kana-containing media at both temperatures. A comparison of two case groups regarding colony counting and OD 600 results did not show any statistically significant association among incubation temperature or ZFN gene targeting efficiency.
    Ptz57r, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 109 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ptz57r/product/Thermo Fisher
    Average 87 stars, based on 109 article reviews
    Price from $9.99 to $1999.99
    ptz57r - by Bioz Stars, 2020-02
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    99
    Thermo Fisher ptz57r t
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    Ptz57r T, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 695 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ptz57r t/product/Thermo Fisher
    Average 99 stars, based on 695 article reviews
    Price from $9.99 to $1999.99
    ptz57r t - by Bioz Stars, 2020-02
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    96
    Promega ptz57r t vector
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    Ptz57r T Vector, supplied by Promega, used in various techniques. Bioz Stars score: 96/100, based on 81 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ptz57r t vector/product/Promega
    Average 96 stars, based on 81 article reviews
    Price from $9.99 to $1999.99
    ptz57r t vector - by Bioz Stars, 2020-02
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    79
    Thermo Fisher ptz57r ta
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    Ptz57r Ta, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ptz57r ta/product/Thermo Fisher
    Average 79 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    ptz57r ta - by Bioz Stars, 2020-02
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    80
    Fisher Scientific vector ptz57r t
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    Vector Ptz57r T, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 80/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vector ptz57r t/product/Fisher Scientific
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    78
    Thermo Fisher instaclone pcr cloning kits ptz57r
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    Instaclone Pcr Cloning Kits Ptz57r, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 78 stars, based on 10 article reviews
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    87
    Thermo Fisher ptz57r t easy vector
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    Ptz57r T Easy Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ptz57r t easy vector/product/Thermo Fisher
    Average 87 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    ptz57r t easy vector - by Bioz Stars, 2020-02
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    79
    Qiagen ptz57r t ta cloning vector
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    Ptz57r T Ta Cloning Vector, supplied by Qiagen, used in various techniques. Bioz Stars score: 79/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ptz57r t ta cloning vector/product/Qiagen
    Average 79 stars, based on 16 article reviews
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    ptz57r t ta cloning vector - by Bioz Stars, 2020-02
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    77
    Thermo Fisher 2886 bp ptz57r t vector
    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid <t>pTZ57R/T-bait-CMV-Neo.</t> CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.
    2886 Bp Ptz57r T Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/2886 bp ptz57r t vector/product/Thermo Fisher
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    78
    Thermo Fisher t overhang ptz57r t plasmid
    Schematic representation of generation of full-length podhfr-ts using overlap extension PCR technique and analysis of amplicons. Four PCR reactions were required to generate the complete sequence: (1) pojr-ts (1134 bp) was PCR amplified from P. ovale DNA and ligated into <t>pTZ57R/T;</t> (2) podhfr-jr (863 bp) and pojr-ts (1134 bp) was acquired using pGEMT- podhfr and pTZ57R/T- pojr-ts as template respectively; (3) and (4) podhfr-ts (1920 bp) was obtained by 2-steps PCR overlap extension, in which dhfr-jr and jr-ts contain overlapping sequences that are complementary for extension shown by half arrow heads indicating the direction in which each fragment can act as primers for DNA polymerase. Dashed lines mark newly synthesized sequence or strand. Horizontal arrows represent primers used in PCR reactions: i, ii, iii, iv, and v are 5′Po-link, 3′PoTS, 5′PoDHFR, 3′Po-link, and 5′NhePo, respectively.
    T Overhang Ptz57r T Plasmid, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t overhang ptz57r t plasmid/product/Thermo Fisher
    Average 78 stars, based on 9 article reviews
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    79
    Thermo Fisher ptz57r t fermentas expression vector
    Schematic representation of generation of full-length podhfr-ts using overlap extension PCR technique and analysis of amplicons. Four PCR reactions were required to generate the complete sequence: (1) pojr-ts (1134 bp) was PCR amplified from P. ovale DNA and ligated into <t>pTZ57R/T;</t> (2) podhfr-jr (863 bp) and pojr-ts (1134 bp) was acquired using pGEMT- podhfr and pTZ57R/T- pojr-ts as template respectively; (3) and (4) podhfr-ts (1920 bp) was obtained by 2-steps PCR overlap extension, in which dhfr-jr and jr-ts contain overlapping sequences that are complementary for extension shown by half arrow heads indicating the direction in which each fragment can act as primers for DNA polymerase. Dashed lines mark newly synthesized sequence or strand. Horizontal arrows represent primers used in PCR reactions: i, ii, iii, iv, and v are 5′Po-link, 3′PoTS, 5′PoDHFR, 3′Po-link, and 5′NhePo, respectively.
    Ptz57r T Fermentas Expression Vector, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ptz57r t fermentas expression vector/product/Thermo Fisher
    Average 79 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    ptz57r t fermentas expression vector - by Bioz Stars, 2020-02
    79/100 stars
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    79
    Macrogen recombinant ptz57r t cyp 141 plasmid
    Schematic representation of generation of full-length podhfr-ts using overlap extension PCR technique and analysis of amplicons. Four PCR reactions were required to generate the complete sequence: (1) pojr-ts (1134 bp) was PCR amplified from P. ovale DNA and ligated into <t>pTZ57R/T;</t> (2) podhfr-jr (863 bp) and pojr-ts (1134 bp) was acquired using pGEMT- podhfr and pTZ57R/T- pojr-ts as template respectively; (3) and (4) podhfr-ts (1920 bp) was obtained by 2-steps PCR overlap extension, in which dhfr-jr and jr-ts contain overlapping sequences that are complementary for extension shown by half arrow heads indicating the direction in which each fragment can act as primers for DNA polymerase. Dashed lines mark newly synthesized sequence or strand. Horizontal arrows represent primers used in PCR reactions: i, ii, iii, iv, and v are 5′Po-link, 3′PoTS, 5′PoDHFR, 3′Po-link, and 5′NhePo, respectively.
    Recombinant Ptz57r T Cyp 141 Plasmid, supplied by Macrogen, used in various techniques. Bioz Stars score: 79/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/recombinant ptz57r t cyp 141 plasmid/product/Macrogen
    Average 79 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    recombinant ptz57r t cyp 141 plasmid - by Bioz Stars, 2020-02
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    Image Search Results


    Map of PTZ 57 R/T-VP7 clone (designed with snap gene software): Sequence analysis indicates that the PTZ57 R/T-VP7 Clone had a full-length copy of segment 9 (VP7) of human rotavirus, and showed 99% homology with RV4 strain (GenBank Sequence ID: M64666.1). The  Bstu-1  restriction enzyme cleaves the PTZ57 R/T-VP7 Clone in the multiple positions. Two of these positions resulted in 635 and 1733 fragments that ultimately released the full-length of VP7 sequence from the plasmid

    Journal: Iranian Journal of Microbiology

    Article Title: An in-vitro transcription assay for development of Rotavirus VP7

    doi:

    Figure Lengend Snippet: Map of PTZ 57 R/T-VP7 clone (designed with snap gene software): Sequence analysis indicates that the PTZ57 R/T-VP7 Clone had a full-length copy of segment 9 (VP7) of human rotavirus, and showed 99% homology with RV4 strain (GenBank Sequence ID: M64666.1). The Bstu-1 restriction enzyme cleaves the PTZ57 R/T-VP7 Clone in the multiple positions. Two of these positions resulted in 635 and 1733 fragments that ultimately released the full-length of VP7 sequence from the plasmid

    Article Snippet: The purified PCR product with 1:3 ratio of 100 ng of 2886 bp linearized vector PTZ57 R/T (Fermentas, Lithuania) and 112.7 ng of 1085 bp PCR product was subcloned into vector.

    Techniques: Software, Sequencing, Plasmid Preparation

    Southern blot and PCR analysis of P. coccineus genomic DNA. (A) Structure of PcPCNA1 and PcPCNA-like1 genes. The nucleotide sequences were analysed using WebGene software. Exons (E), introns (I), and 5′-UTR and 3′-UTR regions are marked. The border sequences of introns termini are placed in the boxes. The positions of internal sites recognized by restriction enzymes used in the Southern blotting analysis are marked. (B) Southern blotting results. 30 μg of the genomic DNA isolated from P. coccineus seedlings were digested with Bam HI (lanes 2 and 8), Bgl II (lanes 3 and 9), Eco RI (lanes 4 and 10), Hin dIII (lanes 5 and 11) or Xba I (lanes 6 and 12), separated in 0.8% agarose gel and subjected to Southern blot procedure with the Pc PCNA1 (lanes 2–6) or Pc PCNA-like1 (lanes 8–12) probe. Lanes 1 and 7: DNA molecular weight marker II Digoxigenin-labelled (Roche). Stars indicate position of DNA fragments detected with Pc PCNA probes. (C) PCR results. PCR was performed using degenerate primers and gDNA isolated from P. coccineus seedlings (lane 3), or plasmid pTZ57R\T DNA containing genomic sequence of the PcPCNA1 gene (lane 1) or of the PcPCNA-like1 gene (lane 2). In negative control, DNA template was omitted (lane 4). Lane M: DNA size standards (1 kb DNA ladder).

    Journal: Journal of Experimental Botany

    Article Title: Identification and functional analysis of PCNA1 and PCNA-like1 genes of Phaseolus coccineus

    doi: 10.1093/jxb/erp354

    Figure Lengend Snippet: Southern blot and PCR analysis of P. coccineus genomic DNA. (A) Structure of PcPCNA1 and PcPCNA-like1 genes. The nucleotide sequences were analysed using WebGene software. Exons (E), introns (I), and 5′-UTR and 3′-UTR regions are marked. The border sequences of introns termini are placed in the boxes. The positions of internal sites recognized by restriction enzymes used in the Southern blotting analysis are marked. (B) Southern blotting results. 30 μg of the genomic DNA isolated from P. coccineus seedlings were digested with Bam HI (lanes 2 and 8), Bgl II (lanes 3 and 9), Eco RI (lanes 4 and 10), Hin dIII (lanes 5 and 11) or Xba I (lanes 6 and 12), separated in 0.8% agarose gel and subjected to Southern blot procedure with the Pc PCNA1 (lanes 2–6) or Pc PCNA-like1 (lanes 8–12) probe. Lanes 1 and 7: DNA molecular weight marker II Digoxigenin-labelled (Roche). Stars indicate position of DNA fragments detected with Pc PCNA probes. (C) PCR results. PCR was performed using degenerate primers and gDNA isolated from P. coccineus seedlings (lane 3), or plasmid pTZ57R\T DNA containing genomic sequence of the PcPCNA1 gene (lane 1) or of the PcPCNA-like1 gene (lane 2). In negative control, DNA template was omitted (lane 4). Lane M: DNA size standards (1 kb DNA ladder).

    Article Snippet: The resulting PCR products were purified and cloned into the pTZ57R\T vector (Fermentas) followed by sequencing.

    Techniques: Southern Blot, Polymerase Chain Reaction, Software, Isolation, Agarose Gel Electrophoresis, Molecular Weight, Marker, Plasmid Preparation, Sequencing, Negative Control

    A; Schematic representation of the pTZ57R/T, B; Schematic representation of the pBAD/gIII A plasmids (obtained from the manufacturers brochure).

    Journal: Research in Pharmaceutical Sciences

    Article Title: Molecualr Cloning of the capsular antigen F1 of Yersinia pestis in pBAD/gIII plasmid

    doi:

    Figure Lengend Snippet: A; Schematic representation of the pTZ57R/T, B; Schematic representation of the pBAD/gIII A plasmids (obtained from the manufacturers brochure).

    Article Snippet: For T/A cloning, pTZ57R/T plasmid ( ; Thermo-scientific, USA) was used.

    Techniques:

    The construction method of Os_GH-pmKate2-N mammalian expression vector. The Os_GH-pTZ57R/T cloning vector and pmKate2-N expression vector were digested with SacI and SacII restriction enzymes to make sticky ends in both Os_GH cDNA fragment and pmKate2-N vector. The digested Os_GH cDNA fragment was ligated into the digested pmKate2-N vector. Finally, the constructed Os_GH-pmKate2-N vector was transformed into DH10B cells for amplification (adapted by authors).

    Journal: Journal of Genetic Engineering & Biotechnology

    Article Title: Construction of ovine GH-pmKate2N expression vector and its uptake by ovine spermatozoa using different methods

    doi: 10.1016/j.jgeb.2017.04.001

    Figure Lengend Snippet: The construction method of Os_GH-pmKate2-N mammalian expression vector. The Os_GH-pTZ57R/T cloning vector and pmKate2-N expression vector were digested with SacI and SacII restriction enzymes to make sticky ends in both Os_GH cDNA fragment and pmKate2-N vector. The digested Os_GH cDNA fragment was ligated into the digested pmKate2-N vector. Finally, the constructed Os_GH-pmKate2-N vector was transformed into DH10B cells for amplification (adapted by authors).

    Article Snippet: 2.3 Cloning and expression vectors construction The visualized band of Os_GH cDNA was extracted from gel using GeneElute™ Gel Extraction Kit (Fermentas, USA) and cloned into pTZ57R/T cloning vector ( ) according to the kit instructions (2886 bp, Fermentas, USA) as follows: 3 μl of pTZ57R/T vector, 2.5 μl of Os_GH cDNA, 6 μl of 10× ligase buffer, 1 μl T4 DNA ligase and 17.5 μl sterile water in total 30 μl reaction mixture and incubated at 22 °C for 1 h for ligation.

    Techniques: Expressing, Plasmid Preparation, Clone Assay, Construct, Transformation Assay, Amplification

    Graphic Comparison of Colony Counting and OD 600 Measurement Results Between Case and Control Groups The dot plot graph of colony counting results; A, and OD600 measurement results; B from the three transformation procedures performed at two different incubation temperatures: 30 and 37°C. The data shown in the graphs reveal a significant reduction in colony numbers and bacterial growth in the E. coli TOP10 containing pZFN, transformed with pTZ57R, compared to the E. coli TOP10 containing the pP15A, kana R vector transformed with pTZ57R, on amp and amp-kana-containing media at both temperatures. A comparison of two case groups regarding colony counting and OD 600 results did not show any statistically significant association among incubation temperature or ZFN gene targeting efficiency.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Zinc Finger Nuclease: A New Approach to Overcome Beta-Lactam Antibiotic Resistance

    doi: 10.5812/jjm.29384

    Figure Lengend Snippet: Graphic Comparison of Colony Counting and OD 600 Measurement Results Between Case and Control Groups The dot plot graph of colony counting results; A, and OD600 measurement results; B from the three transformation procedures performed at two different incubation temperatures: 30 and 37°C. The data shown in the graphs reveal a significant reduction in colony numbers and bacterial growth in the E. coli TOP10 containing pZFN, transformed with pTZ57R, compared to the E. coli TOP10 containing the pP15A, kana R vector transformed with pTZ57R, on amp and amp-kana-containing media at both temperatures. A comparison of two case groups regarding colony counting and OD 600 results did not show any statistically significant association among incubation temperature or ZFN gene targeting efficiency.

    Article Snippet: However, as there was only one potential target site identified by the CoDA platform in the β-lactamase gene of the pTZ57R plasmid (Val 258-Ala 266), the ZFN target site ( ) was cloned in the pTZ57R (Thermo Scientific, Waltham, MA, USA) into the EcoRI and BamHI restriction sites.

    Techniques: Transformation Assay, Incubation, Plasmid Preparation

    The Transformation Results From Experiment 1 at 37°C Figures A – C, show the transformation products of E. coli TOP10 containing pZFN, transformed with pTZ57R, on amp-kana, kana and amp-Containing Media; figures D – F, Display the Transformation Products of E. coli TOP10 containing the pP15A, kana R vector transformed with pTZ57R on the same media; the number of colonies on amp-kana and amp in the case group significantly decreased compared to the control group. The High rate of bacterial growth on kana-containing media ensures that the transformants did not lose their kana R plasmids.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Zinc Finger Nuclease: A New Approach to Overcome Beta-Lactam Antibiotic Resistance

    doi: 10.5812/jjm.29384

    Figure Lengend Snippet: The Transformation Results From Experiment 1 at 37°C Figures A – C, show the transformation products of E. coli TOP10 containing pZFN, transformed with pTZ57R, on amp-kana, kana and amp-Containing Media; figures D – F, Display the Transformation Products of E. coli TOP10 containing the pP15A, kana R vector transformed with pTZ57R on the same media; the number of colonies on amp-kana and amp in the case group significantly decreased compared to the control group. The High rate of bacterial growth on kana-containing media ensures that the transformants did not lose their kana R plasmids.

    Article Snippet: However, as there was only one potential target site identified by the CoDA platform in the β-lactamase gene of the pTZ57R plasmid (Val 258-Ala 266), the ZFN target site ( ) was cloned in the pTZ57R (Thermo Scientific, Waltham, MA, USA) into the EcoRI and BamHI restriction sites.

    Techniques: Transformation Assay, Plasmid Preparation

    Confirmation of the ZFN Target Site Cloning in pTZ57R Lanes 2–5, reveal amplification of a 130-bp PCR product; Lanes 1 and 6, GeneRuler 100-bp DNA ladder (Thermo Scientific, Waltham, MA, USA)

    Journal: Jundishapur Journal of Microbiology

    Article Title: Zinc Finger Nuclease: A New Approach to Overcome Beta-Lactam Antibiotic Resistance

    doi: 10.5812/jjm.29384

    Figure Lengend Snippet: Confirmation of the ZFN Target Site Cloning in pTZ57R Lanes 2–5, reveal amplification of a 130-bp PCR product; Lanes 1 and 6, GeneRuler 100-bp DNA ladder (Thermo Scientific, Waltham, MA, USA)

    Article Snippet: However, as there was only one potential target site identified by the CoDA platform in the β-lactamase gene of the pTZ57R plasmid (Val 258-Ala 266), the ZFN target site ( ) was cloned in the pTZ57R (Thermo Scientific, Waltham, MA, USA) into the EcoRI and BamHI restriction sites.

    Techniques: Clone Assay, Amplification, Polymerase Chain Reaction

    The ZFN Target Site Identified by the CoDA Platform in the β-lactamase Gene of the pTZ57R Plasmid

    Journal: Jundishapur Journal of Microbiology

    Article Title: Zinc Finger Nuclease: A New Approach to Overcome Beta-Lactam Antibiotic Resistance

    doi: 10.5812/jjm.29384

    Figure Lengend Snippet: The ZFN Target Site Identified by the CoDA Platform in the β-lactamase Gene of the pTZ57R Plasmid

    Article Snippet: However, as there was only one potential target site identified by the CoDA platform in the β-lactamase gene of the pTZ57R plasmid (Val 258-Ala 266), the ZFN target site ( ) was cloned in the pTZ57R (Thermo Scientific, Waltham, MA, USA) into the EcoRI and BamHI restriction sites.

    Techniques: Plasmid Preparation

    The Transformation Results From Experiment 1 at 30°C Figures A and B, show the transformation products of E. coli TOP10 Containing pZFN, Transformed With pTZ57R, as the case group on amp-kana and amp containing media; Figures C and D, show the transformation products of E. coli TOP10 containing pP15A, kana R vector transformed with pTZ57R, as the control group, on amp-kana and amp containing media. The plates were incubated overnight at 30°C. A significantly lower level of bacterial growth in the case group compared to the control group was also observed at this temperature.

    Journal: Jundishapur Journal of Microbiology

    Article Title: Zinc Finger Nuclease: A New Approach to Overcome Beta-Lactam Antibiotic Resistance

    doi: 10.5812/jjm.29384

    Figure Lengend Snippet: The Transformation Results From Experiment 1 at 30°C Figures A and B, show the transformation products of E. coli TOP10 Containing pZFN, Transformed With pTZ57R, as the case group on amp-kana and amp containing media; Figures C and D, show the transformation products of E. coli TOP10 containing pP15A, kana R vector transformed with pTZ57R, as the control group, on amp-kana and amp containing media. The plates were incubated overnight at 30°C. A significantly lower level of bacterial growth in the case group compared to the control group was also observed at this temperature.

    Article Snippet: However, as there was only one potential target site identified by the CoDA platform in the β-lactamase gene of the pTZ57R plasmid (Val 258-Ala 266), the ZFN target site ( ) was cloned in the pTZ57R (Thermo Scientific, Waltham, MA, USA) into the EcoRI and BamHI restriction sites.

    Techniques: Transformation Assay, Plasmid Preparation, Incubation

    TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid pTZ57R/T-bait-CMV-Neo. CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.

    Journal: Scientific Reports

    Article Title: TALEN mediated targeted editing of GM2/GD2-synthase gene modulates anchorage independent growth by reducing anoikis resistance in mouse tumor cells

    doi: 10.1038/srep09048

    Figure Lengend Snippet: TALEN mediated targeted integration of CMV-Neomycin cassette into the GM2- synthase locus. Fig. 4a . shows a schematic representation of murine GM2-synthase locus. Blue box indicates the 101 bp genomic region which was amplified to design the donor vector. Pink box with green border (inside the blue box) indicates TALEN target region. Fig. 4b . A schematic of donor plasmid pTZ57R/T-bait-CMV-Neo. CMV-driven neomycin cassette was first cloned into Apa1 site of self-circularized TA cloning vector pTZ57R/T. Then the 101 bp bait sequence was cloned into the C-terminal of neomycin cassette. Yellow box indicates the bait sequence in the donor vector, pink box (inside the yellow box) indicates TALEN binding site. Fig. 4c . A schematic showing NHEJ-mediated integration between TALEN targeted GM2-synthase locus and the donor vector. Junction PCR using primer set (F3 + R4) was used for assessing targeted integration. Blue dotted line indicates the size of PCR products. Fig 4d . A representative agarose gel analysis of PCR genotyping demonstrating targeted integration of neomycin cassette into GM2-synthase locus (all agarose gels were cropped and original images shown in supplementary information . All individual gels were run under same experimental conditions). Genotyping PCR was performed to assess the integration event from Renca-v cells transfected with different amount of donor plasmid with fixed amount of TALEN pair, only donor plasmid or non-transfected cells. Primer pair (F3 + R4) was used to perform the junction PCR, primer pair (F4 + F4) was used to amplify the neomycin cassette and primer pair (F2 + R2) was used to amplify a part of GM2-synthase beyond TALEN target region.

    Article Snippet: High resolution metaphor agarose was purchased from Lonza (Rockland, USA). pTZ57R/T was obtained from Thermo Scientific (EU, Lithuania).

    Techniques: Amplification, Plasmid Preparation, Clone Assay, TA Cloning, Sequencing, Binding Assay, Non-Homologous End Joining, Polymerase Chain Reaction, Agarose Gel Electrophoresis, Transfection

    Schematic representation of generation of full-length podhfr-ts using overlap extension PCR technique and analysis of amplicons. Four PCR reactions were required to generate the complete sequence: (1) pojr-ts (1134 bp) was PCR amplified from P. ovale DNA and ligated into pTZ57R/T; (2) podhfr-jr (863 bp) and pojr-ts (1134 bp) was acquired using pGEMT- podhfr and pTZ57R/T- pojr-ts as template respectively; (3) and (4) podhfr-ts (1920 bp) was obtained by 2-steps PCR overlap extension, in which dhfr-jr and jr-ts contain overlapping sequences that are complementary for extension shown by half arrow heads indicating the direction in which each fragment can act as primers for DNA polymerase. Dashed lines mark newly synthesized sequence or strand. Horizontal arrows represent primers used in PCR reactions: i, ii, iii, iv, and v are 5′Po-link, 3′PoTS, 5′PoDHFR, 3′Po-link, and 5′NhePo, respectively.

    Journal: Parasitology International

    Article Title: Cloning and heterologous expression of Plasmodium ovale dihydrofolate reductase-thymidylate synthase gene

    doi: 10.1016/j.parint.2011.12.004

    Figure Lengend Snippet: Schematic representation of generation of full-length podhfr-ts using overlap extension PCR technique and analysis of amplicons. Four PCR reactions were required to generate the complete sequence: (1) pojr-ts (1134 bp) was PCR amplified from P. ovale DNA and ligated into pTZ57R/T; (2) podhfr-jr (863 bp) and pojr-ts (1134 bp) was acquired using pGEMT- podhfr and pTZ57R/T- pojr-ts as template respectively; (3) and (4) podhfr-ts (1920 bp) was obtained by 2-steps PCR overlap extension, in which dhfr-jr and jr-ts contain overlapping sequences that are complementary for extension shown by half arrow heads indicating the direction in which each fragment can act as primers for DNA polymerase. Dashed lines mark newly synthesized sequence or strand. Horizontal arrows represent primers used in PCR reactions: i, ii, iii, iv, and v are 5′Po-link, 3′PoTS, 5′PoDHFR, 3′Po-link, and 5′NhePo, respectively.

    Article Snippet: The extracted pojr-ts was ligated into T-overhang pTZ57R/T plasmid (Fermentas, Burlington, Canada) and the resulting pTZ57R/T-pojr-ts plasmid construct was used as a template to re-amplify pojr-ts sequence using conditions as described above, except that Pfu polymerase and an elongation time of 2.5 min were used.

    Techniques: Polymerase Chain Reaction, Sequencing, Amplification, Activated Clotting Time Assay, Synthesized