vitro transcription translation ivt ta reactions  (New England Biolabs)


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    New England Biolabs vitro transcription translation ivt ta reactions
    In vitro trials of WNV toehold switches ( A ) Individual in <t>vitro</t> <t>Transcription/Translation</t> <t>(IVT-TA)</t> trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Vitro Transcription Translation Ivt Ta Reactions, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vitro transcription translation ivt ta reactions - by Bioz Stars, 2023-09
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    1) Product Images from "Development of Toehold Switches as a Novel Ribodiagnostic Method for West Nile Virus"

    Article Title: Development of Toehold Switches as a Novel Ribodiagnostic Method for West Nile Virus

    Journal: Genes

    doi: 10.3390/genes14010237

    In vitro trials of WNV toehold switches ( A ) Individual in vitro Transcription/Translation (IVT-TA) trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Figure Legend Snippet: In vitro trials of WNV toehold switches ( A ) Individual in vitro Transcription/Translation (IVT-TA) trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.

    Techniques Used: In Vitro, Isolation

    purexpress in vitro transcription translation kit  (New England Biolabs)


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    New England Biolabs purexpress in vitro transcription translation kit
    Purexpress In Vitro Transcription Translation Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vitro transcription translation ivt ta reactions  (New England Biolabs)


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    New England Biolabs vitro transcription translation ivt ta reactions
    In vitro trials of WNV toehold switches ( A ) Individual in <t>vitro</t> <t>Transcription/Translation</t> <t>(IVT-TA)</t> trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Vitro Transcription Translation Ivt Ta Reactions, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vitro transcription translation ivt ta reactions/product/New England Biolabs
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    vitro transcription translation ivt ta reactions - by Bioz Stars, 2023-09
    96/100 stars

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    1) Product Images from "Development of Toehold Switches as a Novel Ribodiagnostic Method for West Nile Virus"

    Article Title: Development of Toehold Switches as a Novel Ribodiagnostic Method for West Nile Virus

    Journal: Genes

    doi: 10.3390/genes14010237

    In vitro trials of WNV toehold switches ( A ) Individual in vitro Transcription/Translation (IVT-TA) trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Figure Legend Snippet: In vitro trials of WNV toehold switches ( A ) Individual in vitro Transcription/Translation (IVT-TA) trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.

    Techniques Used: In Vitro, Isolation

    vitro cell free transcription translation coupled purexpress system  (New England Biolabs)


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    New England Biolabs vitro cell free transcription translation coupled purexpress system
    Vitro Cell Free Transcription Translation Coupled Purexpress System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vitro transcription translation purexpress system  (New England Biolabs)


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    New England Biolabs vitro transcription translation purexpress system
    Vitro Transcription Translation Purexpress System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    purexpress in vitro transcription translation system  (New England Biolabs)


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    New England Biolabs purexpress in vitro transcription translation system
    Purexpress In Vitro Transcription Translation System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    purexpress cell free transcription translation kit  (New England Biolabs)


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    New England Biolabs purexpress cell free transcription translation kit
    (a) Cells carrying plasmid pBAD18 encoding ShdA II-His6 were grown for 5 hrs in the presence of 0.2 % L-arabinose. Cells were fractionated to produce soluble and membrane samples and analysed by immunoblot with antibodies to the His6 tag, GroEL (cytoplasmic control) and TatA (membrane control). (b-e) In vitro DNAse activity assays using (b) ShdA II 138-524 (c) ShdA I 140-526 (d) ShdA III 135-523 and (e) ShdA IV 135-521 . ShdA proteins and DHFR were synthesised using the cell-free <t>PURExpress</t> kit (NEB). DNAse activity was tested against 10 ng of input DNA. DNA types tested were phage DNA, E. coli MG1655 chromosomal DNA and plasmid (pSG483) DNA. For ShdA I 140-526 , ShdA III 135-523 and ShdA II 138-524 phage DNA was from ϕSipho. For ShdA IV 135-521 phage DNA was from ϕAlma.
    Purexpress Cell Free Transcription Translation Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/purexpress cell free transcription translation kit/product/New England Biolabs
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    Images

    1) Product Images from "Shield co-opts an RmuC domain to mediate phage defence across Pseudomonas species"

    Article Title: Shield co-opts an RmuC domain to mediate phage defence across Pseudomonas species

    Journal: bioRxiv

    doi: 10.1101/2022.11.04.515146

    (a) Cells carrying plasmid pBAD18 encoding ShdA II-His6 were grown for 5 hrs in the presence of 0.2 % L-arabinose. Cells were fractionated to produce soluble and membrane samples and analysed by immunoblot with antibodies to the His6 tag, GroEL (cytoplasmic control) and TatA (membrane control). (b-e) In vitro DNAse activity assays using (b) ShdA II 138-524 (c) ShdA I 140-526 (d) ShdA III 135-523 and (e) ShdA IV 135-521 . ShdA proteins and DHFR were synthesised using the cell-free PURExpress kit (NEB). DNAse activity was tested against 10 ng of input DNA. DNA types tested were phage DNA, E. coli MG1655 chromosomal DNA and plasmid (pSG483) DNA. For ShdA I 140-526 , ShdA III 135-523 and ShdA II 138-524 phage DNA was from ϕSipho. For ShdA IV 135-521 phage DNA was from ϕAlma.
    Figure Legend Snippet: (a) Cells carrying plasmid pBAD18 encoding ShdA II-His6 were grown for 5 hrs in the presence of 0.2 % L-arabinose. Cells were fractionated to produce soluble and membrane samples and analysed by immunoblot with antibodies to the His6 tag, GroEL (cytoplasmic control) and TatA (membrane control). (b-e) In vitro DNAse activity assays using (b) ShdA II 138-524 (c) ShdA I 140-526 (d) ShdA III 135-523 and (e) ShdA IV 135-521 . ShdA proteins and DHFR were synthesised using the cell-free PURExpress kit (NEB). DNAse activity was tested against 10 ng of input DNA. DNA types tested were phage DNA, E. coli MG1655 chromosomal DNA and plasmid (pSG483) DNA. For ShdA I 140-526 , ShdA III 135-523 and ShdA II 138-524 phage DNA was from ϕSipho. For ShdA IV 135-521 phage DNA was from ϕAlma.

    Techniques Used: Plasmid Preparation, Western Blot, In Vitro, Activity Assay

    purexpress in vitro transcription translation system  (New England Biolabs)


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    New England Biolabs purexpress in vitro transcription translation system
    Purexpress In Vitro Transcription Translation System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    purexpress in vitro transcription translation system  (New England Biolabs)


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    New England Biolabs purexpress in vitro transcription translation system
    Purexpress In Vitro Transcription Translation System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    vitro transcription translation purexpress δribosome kit  (New England Biolabs)


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    New England Biolabs vitro transcription translation purexpress δribosome kit
    Vitro Transcription Translation Purexpress δribosome Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    purexpress in vitro transcription translation system  (New England Biolabs)


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    New England Biolabs purexpress in vitro transcription translation system
    Blast hit on P . polymyxa 2020 chromosome with SacB protein from Bacillus subtilis as a query (A). Putative Sac operon from P . polymyxa 2020 (B) and 10–20%SDS PAGE of <t>PURExpress</t> extracts of pSAPv6 (vector), pPpo2020_ sac B (SacB) and pPpo2020_ sac C (SacC). Position of recombinant SacB and SacC proteins indicated by * (C).
    Purexpress In Vitro Transcription Translation System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Production and сharacterization of the exopolysaccharide from strain Paenibacillus polymyxa 2020"

    Article Title: Production and сharacterization of the exopolysaccharide from strain Paenibacillus polymyxa 2020

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0253482

    Blast hit on P . polymyxa 2020 chromosome with SacB protein from Bacillus subtilis as a query (A). Putative Sac operon from P . polymyxa 2020 (B) and 10–20%SDS PAGE of PURExpress extracts of pSAPv6 (vector), pPpo2020_ sac B (SacB) and pPpo2020_ sac C (SacC). Position of recombinant SacB and SacC proteins indicated by * (C).
    Figure Legend Snippet: Blast hit on P . polymyxa 2020 chromosome with SacB protein from Bacillus subtilis as a query (A). Putative Sac operon from P . polymyxa 2020 (B) and 10–20%SDS PAGE of PURExpress extracts of pSAPv6 (vector), pPpo2020_ sac B (SacB) and pPpo2020_ sac C (SacC). Position of recombinant SacB and SacC proteins indicated by * (C).

    Techniques Used: SDS Page, Plasmid Preparation, Recombinant

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    New England Biolabs vitro transcription translation ivt ta reactions
    In vitro trials of WNV toehold switches ( A ) Individual in <t>vitro</t> <t>Transcription/Translation</t> <t>(IVT-TA)</t> trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Vitro Transcription Translation Ivt Ta Reactions, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vitro transcription translation ivt ta reactions/product/New England Biolabs
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    New England Biolabs purexpress in vitro transcription translation kit
    In vitro trials of WNV toehold switches ( A ) Individual in <t>vitro</t> <t>Transcription/Translation</t> <t>(IVT-TA)</t> trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Purexpress In Vitro Transcription Translation Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/purexpress in vitro transcription translation kit/product/New England Biolabs
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    New England Biolabs vitro cell free transcription translation coupled purexpress system
    In vitro trials of WNV toehold switches ( A ) Individual in <t>vitro</t> <t>Transcription/Translation</t> <t>(IVT-TA)</t> trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Vitro Cell Free Transcription Translation Coupled Purexpress System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vitro cell free transcription translation coupled purexpress system/product/New England Biolabs
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    New England Biolabs vitro transcription translation purexpress system
    In vitro trials of WNV toehold switches ( A ) Individual in <t>vitro</t> <t>Transcription/Translation</t> <t>(IVT-TA)</t> trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Vitro Transcription Translation Purexpress System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vitro transcription translation purexpress system/product/New England Biolabs
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    New England Biolabs purexpress in vitro transcription translation system
    In vitro trials of WNV toehold switches ( A ) Individual in <t>vitro</t> <t>Transcription/Translation</t> <t>(IVT-TA)</t> trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.
    Purexpress In Vitro Transcription Translation System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/purexpress in vitro transcription translation system/product/New England Biolabs
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    New England Biolabs purexpress cell free transcription translation kit
    (a) Cells carrying plasmid pBAD18 encoding ShdA II-His6 were grown for 5 hrs in the presence of 0.2 % L-arabinose. Cells were fractionated to produce soluble and membrane samples and analysed by immunoblot with antibodies to the His6 tag, GroEL (cytoplasmic control) and TatA (membrane control). (b-e) In vitro DNAse activity assays using (b) ShdA II 138-524 (c) ShdA I 140-526 (d) ShdA III 135-523 and (e) ShdA IV 135-521 . ShdA proteins and DHFR were synthesised using the cell-free <t>PURExpress</t> kit (NEB). DNAse activity was tested against 10 ng of input DNA. DNA types tested were phage DNA, E. coli MG1655 chromosomal DNA and plasmid (pSG483) DNA. For ShdA I 140-526 , ShdA III 135-523 and ShdA II 138-524 phage DNA was from ϕSipho. For ShdA IV 135-521 phage DNA was from ϕAlma.
    Purexpress Cell Free Transcription Translation Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    New England Biolabs vitro transcription translation purexpress δribosome kit
    (a) Cells carrying plasmid pBAD18 encoding ShdA II-His6 were grown for 5 hrs in the presence of 0.2 % L-arabinose. Cells were fractionated to produce soluble and membrane samples and analysed by immunoblot with antibodies to the His6 tag, GroEL (cytoplasmic control) and TatA (membrane control). (b-e) In vitro DNAse activity assays using (b) ShdA II 138-524 (c) ShdA I 140-526 (d) ShdA III 135-523 and (e) ShdA IV 135-521 . ShdA proteins and DHFR were synthesised using the cell-free <t>PURExpress</t> kit (NEB). DNAse activity was tested against 10 ng of input DNA. DNA types tested were phage DNA, E. coli MG1655 chromosomal DNA and plasmid (pSG483) DNA. For ShdA I 140-526 , ShdA III 135-523 and ShdA II 138-524 phage DNA was from ϕSipho. For ShdA IV 135-521 phage DNA was from ϕAlma.
    Vitro Transcription Translation Purexpress δribosome Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vitro transcription translation purexpress δribosome kit/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    vitro transcription translation purexpress δribosome kit - by Bioz Stars, 2023-09
    96/100 stars
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    In vitro trials of WNV toehold switches ( A ) Individual in vitro Transcription/Translation (IVT-TA) trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.

    Journal: Genes

    Article Title: Development of Toehold Switches as a Novel Ribodiagnostic Method for West Nile Virus

    doi: 10.3390/genes14010237

    Figure Lengend Snippet: In vitro trials of WNV toehold switches ( A ) Individual in vitro Transcription/Translation (IVT-TA) trials of thirty selected GFP-toeholds of WNV. Signal detection was performed in four different channels, corresponding to 507 (blue), 509 (green), 511 (yellow) and 514 (orange) nm across the GFP emission spectrum. Y-axis shows fold enrichment of GFP detection compared to the average of negative controls. Negative controls include IVT-TA trials of different toeholds mixes in the absence of trigger RNA and IVT-TA without any WNV toehold in the presence of 100 ng WNV RNA. ( B ) Independent biological repetitions of the selected toeholds using 100 ng of WNV lineage 2 genomic RNA trigger ( C ) IVT-TA trials of mixed WNV toeholds using both lineage 1 and 2 isolated RNA as trigger. In addition, T10284 that outperformed all other toeholds was also tested individually against both lineage triggers.

    Article Snippet: In vitro Transcription-Translation (IVT-TA) reactions were performed with PURExpress® kit (NEB #E6800L, Bioline Thessaloniki, Greece), as previously described [ ], in the presence of 100ng WNV cultured S/N RNA unless otherwise stated in text.

    Techniques: In Vitro, Isolation

    (a) Cells carrying plasmid pBAD18 encoding ShdA II-His6 were grown for 5 hrs in the presence of 0.2 % L-arabinose. Cells were fractionated to produce soluble and membrane samples and analysed by immunoblot with antibodies to the His6 tag, GroEL (cytoplasmic control) and TatA (membrane control). (b-e) In vitro DNAse activity assays using (b) ShdA II 138-524 (c) ShdA I 140-526 (d) ShdA III 135-523 and (e) ShdA IV 135-521 . ShdA proteins and DHFR were synthesised using the cell-free PURExpress kit (NEB). DNAse activity was tested against 10 ng of input DNA. DNA types tested were phage DNA, E. coli MG1655 chromosomal DNA and plasmid (pSG483) DNA. For ShdA I 140-526 , ShdA III 135-523 and ShdA II 138-524 phage DNA was from ϕSipho. For ShdA IV 135-521 phage DNA was from ϕAlma.

    Journal: bioRxiv

    Article Title: Shield co-opts an RmuC domain to mediate phage defence across Pseudomonas species

    doi: 10.1101/2022.11.04.515146

    Figure Lengend Snippet: (a) Cells carrying plasmid pBAD18 encoding ShdA II-His6 were grown for 5 hrs in the presence of 0.2 % L-arabinose. Cells were fractionated to produce soluble and membrane samples and analysed by immunoblot with antibodies to the His6 tag, GroEL (cytoplasmic control) and TatA (membrane control). (b-e) In vitro DNAse activity assays using (b) ShdA II 138-524 (c) ShdA I 140-526 (d) ShdA III 135-523 and (e) ShdA IV 135-521 . ShdA proteins and DHFR were synthesised using the cell-free PURExpress kit (NEB). DNAse activity was tested against 10 ng of input DNA. DNA types tested were phage DNA, E. coli MG1655 chromosomal DNA and plasmid (pSG483) DNA. For ShdA I 140-526 , ShdA III 135-523 and ShdA II 138-524 phage DNA was from ϕSipho. For ShdA IV 135-521 phage DNA was from ϕAlma.

    Article Snippet: The RmuC domains of ShdA I, ShdA II, ShdA III and ShdA IV homologues were synthesised in vitro using the PURExpress cell-free transcription/translation kit (NEB), from PCR products.

    Techniques: Plasmid Preparation, Western Blot, In Vitro, Activity Assay