top10 escherichia coli transformants  (Thermo Fisher)


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    Structured Review

    Thermo Fisher top10 escherichia coli transformants
    Top10 Escherichia Coli Transformants, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/top10 escherichia coli transformants/product/Thermo Fisher
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    top10 escherichia coli transformants - by Bioz Stars, 2020-07
    85/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Characterization of Bacterial Community Structure in a Drinking Water Distribution System during an Occurrence of Red Water ▿
    Article Snippet: .. The amplification products were purified with a QIAquick PCR cleanup kit (Qiagen, Inc., Chatsworth, CA) and cloned into the TOPO TA cloning vector pCR2.1, with TOP10 Escherichia coli transformants further selected according to the manufacturer's instructions (Invitrogen). .. Cloned inserts were amplified from lysed colonies by PCR with plasmid vector-specific primers M13F and M13R under the same conditions as for the 16S rRNA gene listed above.

    Amplification:

    Article Title: Characterization of Bacterial Community Structure in a Drinking Water Distribution System during an Occurrence of Red Water ▿
    Article Snippet: .. The amplification products were purified with a QIAquick PCR cleanup kit (Qiagen, Inc., Chatsworth, CA) and cloned into the TOPO TA cloning vector pCR2.1, with TOP10 Escherichia coli transformants further selected according to the manufacturer's instructions (Invitrogen). .. Cloned inserts were amplified from lysed colonies by PCR with plasmid vector-specific primers M13F and M13R under the same conditions as for the 16S rRNA gene listed above.

    TA Cloning:

    Article Title: Characterization of Bacterial Community Structure in a Drinking Water Distribution System during an Occurrence of Red Water ▿
    Article Snippet: .. The amplification products were purified with a QIAquick PCR cleanup kit (Qiagen, Inc., Chatsworth, CA) and cloned into the TOPO TA cloning vector pCR2.1, with TOP10 Escherichia coli transformants further selected according to the manufacturer's instructions (Invitrogen). .. Cloned inserts were amplified from lysed colonies by PCR with plasmid vector-specific primers M13F and M13R under the same conditions as for the 16S rRNA gene listed above.

    Purification:

    Article Title: Characterization of Bacterial Community Structure in a Drinking Water Distribution System during an Occurrence of Red Water ▿
    Article Snippet: .. The amplification products were purified with a QIAquick PCR cleanup kit (Qiagen, Inc., Chatsworth, CA) and cloned into the TOPO TA cloning vector pCR2.1, with TOP10 Escherichia coli transformants further selected according to the manufacturer's instructions (Invitrogen). .. Cloned inserts were amplified from lysed colonies by PCR with plasmid vector-specific primers M13F and M13R under the same conditions as for the 16S rRNA gene listed above.

    Polymerase Chain Reaction:

    Article Title: Characterization of Bacterial Community Structure in a Drinking Water Distribution System during an Occurrence of Red Water ▿
    Article Snippet: .. The amplification products were purified with a QIAquick PCR cleanup kit (Qiagen, Inc., Chatsworth, CA) and cloned into the TOPO TA cloning vector pCR2.1, with TOP10 Escherichia coli transformants further selected according to the manufacturer's instructions (Invitrogen). .. Cloned inserts were amplified from lysed colonies by PCR with plasmid vector-specific primers M13F and M13R under the same conditions as for the 16S rRNA gene listed above.

    Plasmid Preparation:

    Article Title: Characterization of Bacterial Community Structure in a Drinking Water Distribution System during an Occurrence of Red Water ▿
    Article Snippet: .. The amplification products were purified with a QIAquick PCR cleanup kit (Qiagen, Inc., Chatsworth, CA) and cloned into the TOPO TA cloning vector pCR2.1, with TOP10 Escherichia coli transformants further selected according to the manufacturer's instructions (Invitrogen). .. Cloned inserts were amplified from lysed colonies by PCR with plasmid vector-specific primers M13F and M13R under the same conditions as for the 16S rRNA gene listed above.

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    Thermo Fisher top10 escherichia coli
    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into <t>TOP10</t> <t>Escherichia</t> coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).
    Top10 Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/top10 escherichia coli/product/Thermo Fisher
    Average 89 stars, based on 25 article reviews
    Price from $9.99 to $1999.99
    top10 escherichia coli - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    84
    Thermo Fisher multishot flexplate top10 competent cells
    Analysis of the pop transcriptional unit. (A) Promoter activity assay for the promoter of pop , which was introduced in the promoterless GFP vector pProbe-NT. Mean fluorescence units of E. coli <t>Top10</t> cells with the different constructs in culture conditions as indicated are given. Error bars show standard deviations. Statistical significance between empty vector (grey bars) and the promoter construct (blue bars) and between growth conditions was tested with a Welch two sample t-test (**/++ p≤0.01; *** p≤0.001; ns, not significant). (B) Test for mono- or polycistronic mRNA. An agarose gel of RT-PCRs is shown. Two different forward primers, binding within ycbG (no. 23) or within pop (no. 24), were combined with a pop reverse primer (no. 25). L: 100 bp DNA Ladder (NEB); 23: PCR with primers 23+25; 24: PCR with primers 24+25. (C) Test for the predicted rho-independent terminator. An agarose gel of RT-PCRs is shown. Two different reverse primers, binding upstream (no. 27) or downstream (no. 28) of the stem loop structure, were combined with a pop forward primer (no. 26). L, 1 kb DNA Ladder (NEB); 27, PCR with primers 27+26; 28, PCR with primers 28+26; d. ORFs, downstream ORFs.
    Multishot Flexplate Top10 Competent Cells, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/multishot flexplate top10 competent cells/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    multishot flexplate top10 competent cells - by Bioz Stars, 2020-07
    84/100 stars
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    92
    Thermo Fisher calcium competent top10 escherichia coli
    Analysis of the pop transcriptional unit. (A) Promoter activity assay for the promoter of pop , which was introduced in the promoterless GFP vector pProbe-NT. Mean fluorescence units of E. coli <t>Top10</t> cells with the different constructs in culture conditions as indicated are given. Error bars show standard deviations. Statistical significance between empty vector (grey bars) and the promoter construct (blue bars) and between growth conditions was tested with a Welch two sample t-test (**/++ p≤0.01; *** p≤0.001; ns, not significant). (B) Test for mono- or polycistronic mRNA. An agarose gel of RT-PCRs is shown. Two different forward primers, binding within ycbG (no. 23) or within pop (no. 24), were combined with a pop reverse primer (no. 25). L: 100 bp DNA Ladder (NEB); 23: PCR with primers 23+25; 24: PCR with primers 24+25. (C) Test for the predicted rho-independent terminator. An agarose gel of RT-PCRs is shown. Two different reverse primers, binding upstream (no. 27) or downstream (no. 28) of the stem loop structure, were combined with a pop forward primer (no. 26). L, 1 kb DNA Ladder (NEB); 27, PCR with primers 27+26; 28, PCR with primers 28+26; d. ORFs, downstream ORFs.
    Calcium Competent Top10 Escherichia Coli, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/calcium competent top10 escherichia coli/product/Thermo Fisher
    Average 92 stars, based on 2 article reviews
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    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Journal: Clinical and Experimental Immunology

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer

    doi: 10.1111/cei.13183

    Figure Lengend Snippet: DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Article Snippet: Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol.

    Techniques: DNA Methylation Assay, Sequencing, Cell Culture, Recombinant, In Vitro, Concentration Assay, Incubation, Flow Cytometry, Cytometry, Expressing, Clone Assay, Plasmid Preparation, Transformation Assay

    DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Journal: Clinical and Experimental Immunology

    Article Title: Tissue‐resident memory T cells are epigenetically cytotoxic with signs of exhaustion in human urinary bladder cancer

    doi: 10.1111/cei.13183

    Figure Lengend Snippet: DNA methylation in the enhancer region regulates perforin transcription. (a) Promoter region sequence containing enhancer in the PRF1 gene was aligned between human and mouse using the VISTA tool. The conservation degree is indicated by the peaks, with coloured peaks (blue for exon and light red for the non‐coding sequence) indicating > 70% conservation. Ten cytosine–phosphate–guanine sites (CpGs) in the enhancer are displayed upstream from the transcription start site (TSS). (b) Sorted CD8 + T cells from healthy donors were cultured initially in the presence of 200 IU/ml recombinant human interleukin (IL)‐2, 5 μg/ml plate‐coated anti‐CD3 and 1 μg/ml anti‐CD28 stimulating antibodies in vitro . 5‐Azacytidine (5‐aza) was added to the culture at a final concentration of 2·5 μM/ml for 48 h, followed by culture medium replacement and incubation for another 48 h. Flow cytometry data demonstrating perforin expression pre‐ and post‐treatment are shown. (c) Perforin – and perforin + CD8 + T cells were sorted. DNA was extracted and bisulfite‐converted from each cell subset, TA‐cloned to pCR4‐TOPO vector and transformed into TOP10 Escherichia coli . Colonies were picked from each cell subset and their DNA was sequenced to measure DNA methylation status of the 10 CpGs in the enhancer. The data display rows represent individually sequenced clone. (d) The bar graphs display total DNA methylation percentage in each CpG from clones in (c).

    Article Snippet: Next, the PCR amplicons were TA‐cloned into pCR4‐TOPO vector (ThermoFisher) and transformed into TOP10 Escherichia coli (ThermoFisher) by heat shock, according to the manufacturer’s protocol.

    Techniques: DNA Methylation Assay, Sequencing, Cell Culture, Recombinant, In Vitro, Concentration Assay, Incubation, Flow Cytometry, Cytometry, Expressing, Clone Assay, Plasmid Preparation, Transformation Assay

    Analysis of the pop transcriptional unit. (A) Promoter activity assay for the promoter of pop , which was introduced in the promoterless GFP vector pProbe-NT. Mean fluorescence units of E. coli Top10 cells with the different constructs in culture conditions as indicated are given. Error bars show standard deviations. Statistical significance between empty vector (grey bars) and the promoter construct (blue bars) and between growth conditions was tested with a Welch two sample t-test (**/++ p≤0.01; *** p≤0.001; ns, not significant). (B) Test for mono- or polycistronic mRNA. An agarose gel of RT-PCRs is shown. Two different forward primers, binding within ycbG (no. 23) or within pop (no. 24), were combined with a pop reverse primer (no. 25). L: 100 bp DNA Ladder (NEB); 23: PCR with primers 23+25; 24: PCR with primers 24+25. (C) Test for the predicted rho-independent terminator. An agarose gel of RT-PCRs is shown. Two different reverse primers, binding upstream (no. 27) or downstream (no. 28) of the stem loop structure, were combined with a pop forward primer (no. 26). L, 1 kb DNA Ladder (NEB); 27, PCR with primers 27+26; 28, PCR with primers 28+26; d. ORFs, downstream ORFs.

    Journal: bioRxiv

    Article Title: A novel pH-regulated, unusual 603 bp overlapping protein coding gene pop is encoded antisense to ompA in Escherichia coli O157:H7 (EHEC)

    doi: 10.1101/852251

    Figure Lengend Snippet: Analysis of the pop transcriptional unit. (A) Promoter activity assay for the promoter of pop , which was introduced in the promoterless GFP vector pProbe-NT. Mean fluorescence units of E. coli Top10 cells with the different constructs in culture conditions as indicated are given. Error bars show standard deviations. Statistical significance between empty vector (grey bars) and the promoter construct (blue bars) and between growth conditions was tested with a Welch two sample t-test (**/++ p≤0.01; *** p≤0.001; ns, not significant). (B) Test for mono- or polycistronic mRNA. An agarose gel of RT-PCRs is shown. Two different forward primers, binding within ycbG (no. 23) or within pop (no. 24), were combined with a pop reverse primer (no. 25). L: 100 bp DNA Ladder (NEB); 23: PCR with primers 23+25; 24: PCR with primers 24+25. (C) Test for the predicted rho-independent terminator. An agarose gel of RT-PCRs is shown. Two different reverse primers, binding upstream (no. 27) or downstream (no. 28) of the stem loop structure, were combined with a pop forward primer (no. 26). L, 1 kb DNA Ladder (NEB); 27, PCR with primers 27+26; 28, PCR with primers 28+26; d. ORFs, downstream ORFs.

    Article Snippet: Vector constructs were transformed in E. coli Top10 cells and plated on LB with required antibiotics.

    Techniques: Activity Assay, Plasmid Preparation, Fluorescence, Construct, Agarose Gel Electrophoresis, Binding Assay, Polymerase Chain Reaction