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  • 94
    Thermo Fisher top10 electrocomp kit
    Top10 Electrocomp Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher top10 transformation
    Top10 Transformation, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher coli top10 transformed bacteria
    Coli Top10 Transformed Bacteria, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher oneshot top10 transformation protocol
    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli <t>Top10</t> cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    Oneshot Top10 Transformation Protocol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 319 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Thermo Fisher transformation escherichia coli strain top10
    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli <t>Top10</t> cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    Transformation Escherichia Coli Strain Top10, supplied by Thermo Fisher, 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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    Thermo Fisher top10 escherichia coli transformants
    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli <t>Top10</t> cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    Top10 Escherichia Coli Transformants, supplied by Thermo Fisher, 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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    94
    Thermo Fisher transform one shot top10 competent escherichia coli cells
    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli <t>Top10</t> cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    Transform One Shot Top10 Competent Escherichia Coli Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Thermo Fisher one shot top10 e
    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli <t>Top10</t> cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    One Shot Top10 E, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 95/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher multishot top10
    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli <t>Top10</t> cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.
    Multishot Top10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher top10
    Dot blot analysis of E. coli strains expressed rhGH in LB and 4YT mediums. A1: GH 1 mgr/ml , A2: GH 100 μgr/ml , A3: GH 10 μgr/ml , A4: GH 1 μgr/ ml , A5: GH 100 ngr/ml , A6: PBS, A7: control – with out loading, A8: <t>Top10</t> none transformed supernatant, A9: Top10 none transformed cell lysate, A10: Top10 transformed LB T0 supernatant, A11: Top10 transformed LB T0 cell lysate, B1: Top10 transformed LB T5 supernatant, B2: Top10 transformed LB T5 supernatant 1/10 diluted, B3: Top10 transformed LB T5 cell lysate, B4: Top10 transformed LB T5 cell lysate 1/10 diluted, B5: Top10 transformed 4YT T0 supernatant, B6: Top10 transformed 4YT T0 cell lysate, B7: Top10 transformed 4YT T5 supernatant, B8: Top10 transformed 4YT T5 supernatant 1/10 diluted, B9: Top10 transformed 4YT T5 cell lysate, B10: Top10 transformed 4YT T5 cell lysate 1/10 diluted, B11 to C11: Top10 repeat 2, D1 to E1: Top10 repeat 3. E2 to F4: repeat 1 of XL1-blue, F5 to G5: repeat 2, G6 to H6: repeat 3. H7 to I9: repeat 1 of JM109, I10 to J10: repeat 2 and J11 to K11: repeat 3.
    Top10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 4148 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher competent one shot top10 cells
    Dot blot analysis of E. coli strains expressed rhGH in LB and 4YT mediums. A1: GH 1 mgr/ml , A2: GH 100 μgr/ml , A3: GH 10 μgr/ml , A4: GH 1 μgr/ ml , A5: GH 100 ngr/ml , A6: PBS, A7: control – with out loading, A8: <t>Top10</t> none transformed supernatant, A9: Top10 none transformed cell lysate, A10: Top10 transformed LB T0 supernatant, A11: Top10 transformed LB T0 cell lysate, B1: Top10 transformed LB T5 supernatant, B2: Top10 transformed LB T5 supernatant 1/10 diluted, B3: Top10 transformed LB T5 cell lysate, B4: Top10 transformed LB T5 cell lysate 1/10 diluted, B5: Top10 transformed 4YT T0 supernatant, B6: Top10 transformed 4YT T0 cell lysate, B7: Top10 transformed 4YT T5 supernatant, B8: Top10 transformed 4YT T5 supernatant 1/10 diluted, B9: Top10 transformed 4YT T5 cell lysate, B10: Top10 transformed 4YT T5 cell lysate 1/10 diluted, B11 to C11: Top10 repeat 2, D1 to E1: Top10 repeat 3. E2 to F4: repeat 1 of XL1-blue, F5 to G5: repeat 2, G6 to H6: repeat 3. H7 to I9: repeat 1 of JM109, I10 to J10: repeat 2 and J11 to K11: repeat 3.
    Competent One Shot Top10 Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 597 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli Top10 cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.

    Journal: PLoS ONE

    Article Title: AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach

    doi: 10.1371/journal.pone.0137652

    Figure Lengend Snippet: AQUA Cloning conditions. (a) The DNA fragment encoding the red fluorescent protein mCherry was PCR amplified with flanking extensions of 16, 24, or 32 bp of homologous sequence overlaps to an SV40 promoter-driven mammalian expression vector which was digested, or PCR amplified. AQUA Cloning was performed with pre-incubations on ice, at room temperature (RT), or at 50°C. DNA mixtures were transformed into chemically competent E . coli Top10 cells and colonies were obtained the next day. (b) Typical numbers of colony forming units (CFU) obtained from each condition. The highest number of CFU was derived from a pre-incubation at room temperature and with 32 bp of shared homology with PCR originating DNA fragments (arrow). The accuracies for each condition were determined by analytical colony PCR from eight selected clones and were extrapolated to the total number of CFU (grey). Abbreviations: P SV40 , simian virus 40 early promoter.

    Article Snippet: E . coli strains The following commercially available strains of E . coli were also used for AQUA Cloning: One Shot TOP10 (Invitrogen cat. No. C4040-03, competency: > 109 CFU/μg), NEB5α (NEB, cat. No. C2987I, competency: 1–3 x 109 CFU/μg), NEB10β (NEB, cat. No. C3019I, competency: 1–3 x 109 CFU/μg), BL21 (DE3) (NEB, cat. No. C2527I, competency: 1–5 x 107 CFU/μg), JM109 (Promega, cat. No. L2005, competency: 108 CFU/μg).

    Techniques: Clone Assay, Polymerase Chain Reaction, Amplification, Sequencing, Expressing, Plasmid Preparation, Transformation Assay, Derivative Assay, Incubation

    AQUA Expression—combined cloning and protein expression. (a) Timeline for AQUA Expression. Cloning and production of recombinant protein in E . coli may be performed within 24 h starting with the PCR until the bacteria are harvested the next day. (b) A 3-DNA fragment cloning was performed by inserting the coding sequence for the red fluorescent protein mCherry into a bacterial T7 promoter-driven expression vector. The vector was split into two parts within the resistance gene for the antibiotic spectinomycin. Therefore, only correctly assembled fragments allow cell growth. (c) AQUA Expression in the expression strain BL21 (DE3) results in red colored bacteria due to mCherry protein production, while the TOP10 strain—lacking the required T7 RNA polymerase—remains colorless.

    Journal: PLoS ONE

    Article Title: AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach

    doi: 10.1371/journal.pone.0137652

    Figure Lengend Snippet: AQUA Expression—combined cloning and protein expression. (a) Timeline for AQUA Expression. Cloning and production of recombinant protein in E . coli may be performed within 24 h starting with the PCR until the bacteria are harvested the next day. (b) A 3-DNA fragment cloning was performed by inserting the coding sequence for the red fluorescent protein mCherry into a bacterial T7 promoter-driven expression vector. The vector was split into two parts within the resistance gene for the antibiotic spectinomycin. Therefore, only correctly assembled fragments allow cell growth. (c) AQUA Expression in the expression strain BL21 (DE3) results in red colored bacteria due to mCherry protein production, while the TOP10 strain—lacking the required T7 RNA polymerase—remains colorless.

    Article Snippet: E . coli strains The following commercially available strains of E . coli were also used for AQUA Cloning: One Shot TOP10 (Invitrogen cat. No. C4040-03, competency: > 109 CFU/μg), NEB5α (NEB, cat. No. C2987I, competency: 1–3 x 109 CFU/μg), NEB10β (NEB, cat. No. C3019I, competency: 1–3 x 109 CFU/μg), BL21 (DE3) (NEB, cat. No. C2527I, competency: 1–5 x 107 CFU/μg), JM109 (Promega, cat. No. L2005, competency: 108 CFU/μg).

    Techniques: Expressing, Clone Assay, Recombinant, Polymerase Chain Reaction, Sequencing, Plasmid Preparation

    AQUA Cloning: a dvanced qu ick a ssembly cloning. (a) DNA parts are produced by PCR, or restriction digest (or both). Oligonucleotides are designed to contribute flanking homologous regions to adjacent DNA fragments of optimally 32 bp in length. DNA parts are assembled into a circular plasmid by sequence-determined directionality. (b) AQUA Cloning work-flow. (1) DNA parts are generated by PCR amplification, or derived from an enzymatic digestion. (2) Next, DNA parts are purified by gel-electrophoresis and (3) mixed and simply incubated in water prior to transformation into chemically competent E . coli Top10 cells for in vivo assembly. (4) Finally, obtained colonies are confirmed for correct assembly by standard methods such as analytical PCR, restriction digest, or comprehensive sequencing.

    Journal: PLoS ONE

    Article Title: AQUA Cloning: A Versatile and Simple Enzyme-Free Cloning Approach

    doi: 10.1371/journal.pone.0137652

    Figure Lengend Snippet: AQUA Cloning: a dvanced qu ick a ssembly cloning. (a) DNA parts are produced by PCR, or restriction digest (or both). Oligonucleotides are designed to contribute flanking homologous regions to adjacent DNA fragments of optimally 32 bp in length. DNA parts are assembled into a circular plasmid by sequence-determined directionality. (b) AQUA Cloning work-flow. (1) DNA parts are generated by PCR amplification, or derived from an enzymatic digestion. (2) Next, DNA parts are purified by gel-electrophoresis and (3) mixed and simply incubated in water prior to transformation into chemically competent E . coli Top10 cells for in vivo assembly. (4) Finally, obtained colonies are confirmed for correct assembly by standard methods such as analytical PCR, restriction digest, or comprehensive sequencing.

    Article Snippet: E . coli strains The following commercially available strains of E . coli were also used for AQUA Cloning: One Shot TOP10 (Invitrogen cat. No. C4040-03, competency: > 109 CFU/μg), NEB5α (NEB, cat. No. C2987I, competency: 1–3 x 109 CFU/μg), NEB10β (NEB, cat. No. C3019I, competency: 1–3 x 109 CFU/μg), BL21 (DE3) (NEB, cat. No. C2527I, competency: 1–5 x 107 CFU/μg), JM109 (Promega, cat. No. L2005, competency: 108 CFU/μg).

    Techniques: Clone Assay, Produced, Polymerase Chain Reaction, Plasmid Preparation, Sequencing, Flow Cytometry, Generated, Amplification, Derivative Assay, Purification, Nucleic Acid Electrophoresis, Incubation, Transformation Assay, In Vivo

    Dot blot analysis of E. coli strains expressed rhGH in LB and 4YT mediums. A1: GH 1 mgr/ml , A2: GH 100 μgr/ml , A3: GH 10 μgr/ml , A4: GH 1 μgr/ ml , A5: GH 100 ngr/ml , A6: PBS, A7: control – with out loading, A8: Top10 none transformed supernatant, A9: Top10 none transformed cell lysate, A10: Top10 transformed LB T0 supernatant, A11: Top10 transformed LB T0 cell lysate, B1: Top10 transformed LB T5 supernatant, B2: Top10 transformed LB T5 supernatant 1/10 diluted, B3: Top10 transformed LB T5 cell lysate, B4: Top10 transformed LB T5 cell lysate 1/10 diluted, B5: Top10 transformed 4YT T0 supernatant, B6: Top10 transformed 4YT T0 cell lysate, B7: Top10 transformed 4YT T5 supernatant, B8: Top10 transformed 4YT T5 supernatant 1/10 diluted, B9: Top10 transformed 4YT T5 cell lysate, B10: Top10 transformed 4YT T5 cell lysate 1/10 diluted, B11 to C11: Top10 repeat 2, D1 to E1: Top10 repeat 3. E2 to F4: repeat 1 of XL1-blue, F5 to G5: repeat 2, G6 to H6: repeat 3. H7 to I9: repeat 1 of JM109, I10 to J10: repeat 2 and J11 to K11: repeat 3.

    Journal: Journal of Research in Medical Sciences : The Official Journal of Isfahan University of Medical Sciences

    Article Title: Optimization of production of recombinant human growth hormone in Escherichia coli

    doi:

    Figure Lengend Snippet: Dot blot analysis of E. coli strains expressed rhGH in LB and 4YT mediums. A1: GH 1 mgr/ml , A2: GH 100 μgr/ml , A3: GH 10 μgr/ml , A4: GH 1 μgr/ ml , A5: GH 100 ngr/ml , A6: PBS, A7: control – with out loading, A8: Top10 none transformed supernatant, A9: Top10 none transformed cell lysate, A10: Top10 transformed LB T0 supernatant, A11: Top10 transformed LB T0 cell lysate, B1: Top10 transformed LB T5 supernatant, B2: Top10 transformed LB T5 supernatant 1/10 diluted, B3: Top10 transformed LB T5 cell lysate, B4: Top10 transformed LB T5 cell lysate 1/10 diluted, B5: Top10 transformed 4YT T0 supernatant, B6: Top10 transformed 4YT T0 cell lysate, B7: Top10 transformed 4YT T5 supernatant, B8: Top10 transformed 4YT T5 supernatant 1/10 diluted, B9: Top10 transformed 4YT T5 cell lysate, B10: Top10 transformed 4YT T5 cell lysate 1/10 diluted, B11 to C11: Top10 repeat 2, D1 to E1: Top10 repeat 3. E2 to F4: repeat 1 of XL1-blue, F5 to G5: repeat 2, G6 to H6: repeat 3. H7 to I9: repeat 1 of JM109, I10 to J10: repeat 2 and J11 to K11: repeat 3.

    Article Snippet: Bacterial Strains and Plasmid The E. coli host strains used in this study were TOP10 (Invitrogen, USA), XL1-blue and JM109 (Cinagen, Iran).

    Techniques: Dot Blot, Transformation Assay

    Comparison of transformation efficiency between different E.coli strains (A) Top10 (B) XL1-blue and (C) JM109

    Journal: Journal of Research in Medical Sciences : The Official Journal of Isfahan University of Medical Sciences

    Article Title: Optimization of production of recombinant human growth hormone in Escherichia coli

    doi:

    Figure Lengend Snippet: Comparison of transformation efficiency between different E.coli strains (A) Top10 (B) XL1-blue and (C) JM109

    Article Snippet: Bacterial Strains and Plasmid The E. coli host strains used in this study were TOP10 (Invitrogen, USA), XL1-blue and JM109 (Cinagen, Iran).

    Techniques: Transformation Assay