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  • 99
    Thermo Fisher filter capped cages
    Filter Capped Cages, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/filter capped cages/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
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    filter capped cages - by Bioz Stars, 2020-08
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    88
    Becton Dickinson capped tubes
    Capped Tubes, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped tubes/product/Becton Dickinson
    Average 88 stars, based on 19 article reviews
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    capped tubes - by Bioz Stars, 2020-08
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    88
    Becton Dickinson capped falcon tubes
    Capped Falcon Tubes, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped falcon tubes/product/Becton Dickinson
    Average 88 stars, based on 9 article reviews
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    capped falcon tubes - by Bioz Stars, 2020-08
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    88
    Fisher Scientific capped microcentrifuge tube
    Capped Microcentrifuge Tube, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 88/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped microcentrifuge tube/product/Fisher Scientific
    Average 88 stars, based on 34 article reviews
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    capped microcentrifuge tube - by Bioz Stars, 2020-08
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    89
    Sarstedt screw capped tubes
    Screw Capped Tubes, supplied by Sarstedt, used in various techniques. Bioz Stars score: 89/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/screw capped tubes/product/Sarstedt
    Average 89 stars, based on 16 article reviews
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    93
    Millipore tannic capped aunps
    Naked eye detection of cancer using <t>AuNPs.</t> a Schematic of the assay and proposed mechanism for different DNA types. b Mean relative absorbance values A 520/658 of 10 nm tannic-capped <t>AuNP</t> (pink) and AuNP- gDNA solution for unmethylated WGA (black), BT474 breast cancer cell line (red), and Aza treated Jurkat (light blue). Inset, the representative coloured solution. TEM images of c AuNPs alone, d AuNP-gDNA solution for unmethylated WGA, e AuNP with BT474 cancer DNA and f AuNP with 100% methylated Jurkat DNA (no salt was added during sample preparation, scale bars are 500 nm for normal images and 100 nm for the zoomed images in the black boxes). g Box plot showing the mean relative absorbance values A 520/658 of AuNP-gDNA solution for cancer and normal cells extracted from breast, prostate and lymph node tissues, Right: The ROC analysis and diagnostic test evaluation shows the Disease Prevalence (DP), positive predictive values (PPV), negative predictive values (NPV) and accuracy of the method. h Box plot showing the mean relative absorbance values A 520/658 of AuNP-gDNA solution for DNA samples derived from plasma samples of breast and colorectal cancer patients or healthy donors, Right: The ROC analysis and diagnostic test evaluation shows the Disease Prevalence (DP), positive predictive values (PPV), negative predictive values (NPV) and accuracy of the method. In the box and whisker plot, the middle lines of the boxes represent the median (50th percentile) and the terminal line of the boxes represents the 25th to 75th percentile. The whiskers represent the lowest and the highest value
    Tannic Capped Aunps, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tannic capped aunps/product/Millipore
    Average 93 stars, based on 6 article reviews
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    91
    TriLink capped rna
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Capped Rna, supplied by TriLink, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped rna/product/TriLink
    Average 91 stars, based on 4 article reviews
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    capped rna - by Bioz Stars, 2020-08
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    89
    Sarstedt capped micro tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Capped Micro Tubes, supplied by Sarstedt, used in various techniques. Bioz Stars score: 89/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped micro tubes/product/Sarstedt
    Average 89 stars, based on 24 article reviews
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    capped micro tubes - by Bioz Stars, 2020-08
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    85
    USA Scientific Inc small capped tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Small Capped Tubes, supplied by USA Scientific Inc, used in various techniques. Bioz Stars score: 85/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/small capped tubes/product/USA Scientific Inc
    Average 85 stars, based on 15 article reviews
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    small capped tubes - by Bioz Stars, 2020-08
    85/100 stars
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    88
    Greiner Bio sterile capped tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Sterile Capped Tubes, supplied by Greiner Bio, used in various techniques. Bioz Stars score: 88/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sterile capped tubes/product/Greiner Bio
    Average 88 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    sterile capped tubes - by Bioz Stars, 2020-08
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    85
    Fisher Scientific capped centrifuge tube
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Capped Centrifuge Tube, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped centrifuge tube/product/Fisher Scientific
    Average 85 stars, based on 4 article reviews
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    capped centrifuge tube - by Bioz Stars, 2020-08
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    93
    Corning Life Sciences filter capped tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Filter Capped Tubes, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/filter capped tubes/product/Corning Life Sciences
    Average 93 stars, based on 5 article reviews
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    filter capped tubes - by Bioz Stars, 2020-08
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    88
    Supelco screw capped vial
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Screw Capped Vial, supplied by Supelco, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/screw capped vial/product/Supelco
    Average 88 stars, based on 9 article reviews
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    screw capped vial - by Bioz Stars, 2020-08
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    88
    Eppendorf AG un capped eppendorf
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Un Capped Eppendorf, supplied by Eppendorf AG, used in various techniques. Bioz Stars score: 88/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/un capped eppendorf/product/Eppendorf AG
    Average 88 stars, based on 12 article reviews
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    un capped eppendorf - by Bioz Stars, 2020-08
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    85
    Becton Dickinson strainer capped tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Strainer Capped Tubes, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/strainer capped tubes/product/Becton Dickinson
    Average 85 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    strainer capped tubes - by Bioz Stars, 2020-08
    85/100 stars
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    85
    Becton Dickinson capped polystyrene tube
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Capped Polystyrene Tube, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped polystyrene tube/product/Becton Dickinson
    Average 85 stars, based on 5 article reviews
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    capped polystyrene tube - by Bioz Stars, 2020-08
    85/100 stars
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    85
    Starna septum capped glass cuvette
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Septum Capped Glass Cuvette, supplied by Starna, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/septum capped glass cuvette/product/Starna
    Average 85 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    septum capped glass cuvette - by Bioz Stars, 2020-08
    85/100 stars
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    89
    Thermo Fisher capped rna
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Capped Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 226 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped rna/product/Thermo Fisher
    Average 89 stars, based on 226 article reviews
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    capped rna - by Bioz Stars, 2020-08
    89/100 stars
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    85
    Becton Dickinson capped polypropylene tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Capped Polypropylene Tubes, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped polypropylene tubes/product/Becton Dickinson
    Average 85 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    capped polypropylene tubes - by Bioz Stars, 2020-08
    85/100 stars
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    88
    Sarstedt capped polystyrene tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Capped Polystyrene Tubes, supplied by Sarstedt, used in various techniques. Bioz Stars score: 88/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/capped polystyrene tubes/product/Sarstedt
    Average 88 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    capped polystyrene tubes - by Bioz Stars, 2020-08
    88/100 stars
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    91
    Biopure Corporation citrate capped biopure
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Citrate Capped Biopure, supplied by Biopure Corporation, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/citrate capped biopure/product/Biopure Corporation
    Average 91 stars, based on 3 article reviews
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    citrate capped biopure - by Bioz Stars, 2020-08
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    85
    Avantor screw capped cryogenic vials
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Screw Capped Cryogenic Vials, supplied by Avantor, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/screw capped cryogenic vials/product/Avantor
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    screw capped cryogenic vials - by Bioz Stars, 2020-08
    85/100 stars
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    85
    Corning Life Sciences pyrex screw capped tubes
    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter <t>5΄</t> and 3΄ ends separately and in competition with <t>RNA–RNA</t> intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D
    Pyrex Screw Capped Tubes, supplied by Corning Life Sciences, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pyrex screw capped tubes/product/Corning Life Sciences
    Average 85 stars, based on 5 article reviews
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    pyrex screw capped tubes - by Bioz Stars, 2020-08
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    Image Search Results


    Naked eye detection of cancer using AuNPs. a Schematic of the assay and proposed mechanism for different DNA types. b Mean relative absorbance values A 520/658 of 10 nm tannic-capped AuNP (pink) and AuNP- gDNA solution for unmethylated WGA (black), BT474 breast cancer cell line (red), and Aza treated Jurkat (light blue). Inset, the representative coloured solution. TEM images of c AuNPs alone, d AuNP-gDNA solution for unmethylated WGA, e AuNP with BT474 cancer DNA and f AuNP with 100% methylated Jurkat DNA (no salt was added during sample preparation, scale bars are 500 nm for normal images and 100 nm for the zoomed images in the black boxes). g Box plot showing the mean relative absorbance values A 520/658 of AuNP-gDNA solution for cancer and normal cells extracted from breast, prostate and lymph node tissues, Right: The ROC analysis and diagnostic test evaluation shows the Disease Prevalence (DP), positive predictive values (PPV), negative predictive values (NPV) and accuracy of the method. h Box plot showing the mean relative absorbance values A 520/658 of AuNP-gDNA solution for DNA samples derived from plasma samples of breast and colorectal cancer patients or healthy donors, Right: The ROC analysis and diagnostic test evaluation shows the Disease Prevalence (DP), positive predictive values (PPV), negative predictive values (NPV) and accuracy of the method. In the box and whisker plot, the middle lines of the boxes represent the median (50th percentile) and the terminal line of the boxes represents the 25th to 75th percentile. The whiskers represent the lowest and the highest value

    Journal: Nature Communications

    Article Title: Epigenetically reprogrammed methylation landscape drives the DNA self-assembly and serves as a universal cancer biomarker

    doi: 10.1038/s41467-018-07214-w

    Figure Lengend Snippet: Naked eye detection of cancer using AuNPs. a Schematic of the assay and proposed mechanism for different DNA types. b Mean relative absorbance values A 520/658 of 10 nm tannic-capped AuNP (pink) and AuNP- gDNA solution for unmethylated WGA (black), BT474 breast cancer cell line (red), and Aza treated Jurkat (light blue). Inset, the representative coloured solution. TEM images of c AuNPs alone, d AuNP-gDNA solution for unmethylated WGA, e AuNP with BT474 cancer DNA and f AuNP with 100% methylated Jurkat DNA (no salt was added during sample preparation, scale bars are 500 nm for normal images and 100 nm for the zoomed images in the black boxes). g Box plot showing the mean relative absorbance values A 520/658 of AuNP-gDNA solution for cancer and normal cells extracted from breast, prostate and lymph node tissues, Right: The ROC analysis and diagnostic test evaluation shows the Disease Prevalence (DP), positive predictive values (PPV), negative predictive values (NPV) and accuracy of the method. h Box plot showing the mean relative absorbance values A 520/658 of AuNP-gDNA solution for DNA samples derived from plasma samples of breast and colorectal cancer patients or healthy donors, Right: The ROC analysis and diagnostic test evaluation shows the Disease Prevalence (DP), positive predictive values (PPV), negative predictive values (NPV) and accuracy of the method. In the box and whisker plot, the middle lines of the boxes represent the median (50th percentile) and the terminal line of the boxes represents the 25th to 75th percentile. The whiskers represent the lowest and the highest value

    Article Snippet: Detection by AuNP system Experiments are performed using 8.5 µL of 10 nm Tannic-capped AuNPs (Sigma), which were mixed with 1 µL of DNA samples (i.e. genomic DNA at 50 ng/µL concentration or cfDNA at 1 pg/µL concentration).

    Techniques: Whole Genome Amplification, Transmission Electron Microscopy, Methylation, Sample Prep, Diagnostic Assay, Derivative Assay, Whisker Assay

    Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter 5΄ and 3΄ ends separately and in competition with RNA–RNA intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D

    Journal: Nucleic Acids Research

    Article Title: An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase

    doi: 10.1093/nar/gkx043

    Figure Lengend Snippet: Interaction of influenza polymerase with the promoter. Influenza polymerase binds the promoter 5΄ and 3΄ ends separately and in competition with RNA–RNA intermolecular interactions. ( A ) Due to partial complementarity, the viral 5΄ and 3΄ RNA ends anneal to form a double-stranded ‘panhandle’ conformation as quantified by a simple FRET assay (v5΄, v3΄/c5΄, c3΄ correspond to the genomic/anti-genomic (complementary) 5΄ and 3΄ RNA ends, respectively; all RNAs are 18 nucleotides each if not indicated otherwise; numbering starts from the 5΄ end or 3΄ end of the 5΄ or 3΄ RNA respectively; see Material and Methods). ( B ) Using a fluorescence polarization based assay, the interaction of influenza polymerase and each promoter RNA was quantified and revealed each RNA separately to interact with the polymerase in a one-to-one stoichiometry and with high affinity ( K D

    Article Snippet: To monitor RNA-primed polymerization, reactions were supplemented by 0.5 mM ApG (TriLink BioTechnologies, San Diego, USA or IBA, Goettingen, Germany) or by 1 μM of capped-RNA (5΄-m7 GpppAAUCUAUAAUAG-3΄; TriLink BioTechnologies, San Diego, USA) if not indicated otherwise.

    Techniques: Fluorescence

    Schematic model of influenza polymerase initiating RNA synthesis at the vRNA promoter. The 5΄-RNA is firmly anchored to the polymerase (beige sphere), the 3΄ template RNA positioned for terminal initiation at the active site by intermolecular interactions of the distal promoter RNAs (5΄-3΄ base-pairing region; highlighted, green base-pairs). Simultaneous binding of ATP at the post-translocation site and GTP at the active site (pre-translocation) followed by formation of the first phosphodiester bond is bypassed by an appropriate primer ending in AG-3΄. CTP, the next nucleotide to be incorporated and directed by G3 on the vRNA template is the first (and within the five ultimate nucleotides the only) distinction from the cRNA template. To allow for translocation of the template RNA and continued RNA synthesis (elongation), the 5΄-3΄ base-pairing region must dissipate which we propose is coupled to the newly emerging base-pairs between product and template RNA downstream the active site (pink). At the cRNA promoter, terminal fast initiation of RNA synthesis is prevented since with UTP directed to position A3, an A–U base pair is formed, instead of the G–C base-pair in the case of vRNA, which apparently does not compensate for breaking the cRNA promoter's 5΄-3΄ base-pairing region. At the expense of rate, the cRNA promoter re-directs RNA synthesis via internal initiation followed by re-align (see Figure 5 ).

    Journal: Nucleic Acids Research

    Article Title: An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase

    doi: 10.1093/nar/gkx043

    Figure Lengend Snippet: Schematic model of influenza polymerase initiating RNA synthesis at the vRNA promoter. The 5΄-RNA is firmly anchored to the polymerase (beige sphere), the 3΄ template RNA positioned for terminal initiation at the active site by intermolecular interactions of the distal promoter RNAs (5΄-3΄ base-pairing region; highlighted, green base-pairs). Simultaneous binding of ATP at the post-translocation site and GTP at the active site (pre-translocation) followed by formation of the first phosphodiester bond is bypassed by an appropriate primer ending in AG-3΄. CTP, the next nucleotide to be incorporated and directed by G3 on the vRNA template is the first (and within the five ultimate nucleotides the only) distinction from the cRNA template. To allow for translocation of the template RNA and continued RNA synthesis (elongation), the 5΄-3΄ base-pairing region must dissipate which we propose is coupled to the newly emerging base-pairs between product and template RNA downstream the active site (pink). At the cRNA promoter, terminal fast initiation of RNA synthesis is prevented since with UTP directed to position A3, an A–U base pair is formed, instead of the G–C base-pair in the case of vRNA, which apparently does not compensate for breaking the cRNA promoter's 5΄-3΄ base-pairing region. At the expense of rate, the cRNA promoter re-directs RNA synthesis via internal initiation followed by re-align (see Figure 5 ).

    Article Snippet: To monitor RNA-primed polymerization, reactions were supplemented by 0.5 mM ApG (TriLink BioTechnologies, San Diego, USA or IBA, Goettingen, Germany) or by 1 μM of capped-RNA (5΄-m7 GpppAAUCUAUAAUAG-3΄; TriLink BioTechnologies, San Diego, USA) if not indicated otherwise.

    Techniques: Binding Assay, Translocation Assay

    Schematic and hypothetical model of internal initiation of RNA synthesis at the cRNA promoter involving prime-and-re-align . Influenza B polymerase is depicted by a beige sphere with the location of the active site indicated by the arrow. The c5΄ end (pink) is firmly anchored to the polymerase, and helps position the c3΄ 1–18 template RNA (pink) for terminal or internal initiation at the active site. The depicted positioning of the c3΄ template RNA for internal initiation would be stabilized by a 5΄-3΄ base-pairing region involving (at least) c5΄ nucleotides G11, A12 and G13 and c3΄ nucleotides C12, U13, C14, in analogy to the vRNA promoter. The initial ATP would bind opposite of U4 at the active site in the pre-translocation conformation. Template RNA translocation by one step places the initial ATP into the post-translocation state, allowing binding of incoming GTP opposite to C5 at the active site and its incorporation to form pppApG. With the template RNA having translocated by 1 position, strain in the 5΄-3΄ base-pairing region would be generated. Further translocation of the template RNA and pppApG to the post-translocation conformation allows binding of incoming UTP opposite of A6 at the active site and its subsequent incorporation. However, this would build-up additional unfavourable strain in the 5΄-3΄ base-pairing region which would need to be relieved. However, breaking the strong 5΄-3΄ base-pairing region formed by the cRNA promoter (2x GC, 1x AU) is not the most favorable outcome at this stage since the newly formed base-pairs between the product RNA and the template RNA (2x AU, 1x GC) downstream of the active site do not compensate energetically. More favorably, the template RNA would translocate three steps backward while the product RNA stays in place. This alternative position of the template would then promote incorporation of the next nucleotide, through binding of ATP opposite of U4, as this restores three unstrained base-pairs in an alternative 5΄-3΄ base-pairing region. Translocation, binding of GTP opposite of C5 and formation of pppApGpUpApG would again build-up strain in the 5΄-3΄ base-pairing region. However, at this stage more newly formed base-pairs between the product RNA and the template RNA are available (compared to having initiated terminally) which would compensate for breaking the 5΄-3΄ base-pairing region and allow RNA synthesis to progress to elongation.

    Journal: Nucleic Acids Research

    Article Title: An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase

    doi: 10.1093/nar/gkx043

    Figure Lengend Snippet: Schematic and hypothetical model of internal initiation of RNA synthesis at the cRNA promoter involving prime-and-re-align . Influenza B polymerase is depicted by a beige sphere with the location of the active site indicated by the arrow. The c5΄ end (pink) is firmly anchored to the polymerase, and helps position the c3΄ 1–18 template RNA (pink) for terminal or internal initiation at the active site. The depicted positioning of the c3΄ template RNA for internal initiation would be stabilized by a 5΄-3΄ base-pairing region involving (at least) c5΄ nucleotides G11, A12 and G13 and c3΄ nucleotides C12, U13, C14, in analogy to the vRNA promoter. The initial ATP would bind opposite of U4 at the active site in the pre-translocation conformation. Template RNA translocation by one step places the initial ATP into the post-translocation state, allowing binding of incoming GTP opposite to C5 at the active site and its incorporation to form pppApG. With the template RNA having translocated by 1 position, strain in the 5΄-3΄ base-pairing region would be generated. Further translocation of the template RNA and pppApG to the post-translocation conformation allows binding of incoming UTP opposite of A6 at the active site and its subsequent incorporation. However, this would build-up additional unfavourable strain in the 5΄-3΄ base-pairing region which would need to be relieved. However, breaking the strong 5΄-3΄ base-pairing region formed by the cRNA promoter (2x GC, 1x AU) is not the most favorable outcome at this stage since the newly formed base-pairs between the product RNA and the template RNA (2x AU, 1x GC) downstream of the active site do not compensate energetically. More favorably, the template RNA would translocate three steps backward while the product RNA stays in place. This alternative position of the template would then promote incorporation of the next nucleotide, through binding of ATP opposite of U4, as this restores three unstrained base-pairs in an alternative 5΄-3΄ base-pairing region. Translocation, binding of GTP opposite of C5 and formation of pppApGpUpApG would again build-up strain in the 5΄-3΄ base-pairing region. However, at this stage more newly formed base-pairs between the product RNA and the template RNA are available (compared to having initiated terminally) which would compensate for breaking the 5΄-3΄ base-pairing region and allow RNA synthesis to progress to elongation.

    Article Snippet: To monitor RNA-primed polymerization, reactions were supplemented by 0.5 mM ApG (TriLink BioTechnologies, San Diego, USA or IBA, Goettingen, Germany) or by 1 μM of capped-RNA (5΄-m7 GpppAAUCUAUAAUAG-3΄; TriLink BioTechnologies, San Diego, USA) if not indicated otherwise.

    Techniques: Translocation Assay, Binding Assay, Generated

    Primed versus unprimed initiation of RNA synthesis by influenza B polymerase in vitro . ( A ) ApG, in a concentration-dependent manner (colored triangles), accelerates unprimed RNA synthesis (black triangles) to the level of capped RNA primed RNA synthesis reaction (black circles). ( B ) Affinities ( K M ) for the primer(s) in accelerating initiation of RNA synthesis show a preference of the capped RNA over non-capped RNA ending in AG-3΄ over minimally ApG. Error bars indicate the standard deviation of the mean of at least duplicate experiments (see Supplementary Figure S8 ). ( C ) Schematic depicting the initiation of RNA synthesis catalysed by influenza B polymerase at the vRNA promotor (v5΄ + v3΄). The 5΄-3΄ base-pairing region (involving v5΄ nt 11–13 and v3΄ nt 10–12) positions the 3΄ template RNA at the polymerase's active site. Formation of the first phosphodiester bond (yielding ApG) is rate-limiting for initiating RNA synthesis and bypassed by supplying an appropriate primer. ApG serves as a minimal primer and accelerates unprimed RNA synthesis by annealing to the U1C2 at the 3΄ template extremity. This ‘primed initiation complex’ (PRIC) conformation is further stabilized by favourable interactions of the polymerase with the primer's RNA-moiety and cap-structure.

    Journal: Nucleic Acids Research

    Article Title: An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase

    doi: 10.1093/nar/gkx043

    Figure Lengend Snippet: Primed versus unprimed initiation of RNA synthesis by influenza B polymerase in vitro . ( A ) ApG, in a concentration-dependent manner (colored triangles), accelerates unprimed RNA synthesis (black triangles) to the level of capped RNA primed RNA synthesis reaction (black circles). ( B ) Affinities ( K M ) for the primer(s) in accelerating initiation of RNA synthesis show a preference of the capped RNA over non-capped RNA ending in AG-3΄ over minimally ApG. Error bars indicate the standard deviation of the mean of at least duplicate experiments (see Supplementary Figure S8 ). ( C ) Schematic depicting the initiation of RNA synthesis catalysed by influenza B polymerase at the vRNA promotor (v5΄ + v3΄). The 5΄-3΄ base-pairing region (involving v5΄ nt 11–13 and v3΄ nt 10–12) positions the 3΄ template RNA at the polymerase's active site. Formation of the first phosphodiester bond (yielding ApG) is rate-limiting for initiating RNA synthesis and bypassed by supplying an appropriate primer. ApG serves as a minimal primer and accelerates unprimed RNA synthesis by annealing to the U1C2 at the 3΄ template extremity. This ‘primed initiation complex’ (PRIC) conformation is further stabilized by favourable interactions of the polymerase with the primer's RNA-moiety and cap-structure.

    Article Snippet: To monitor RNA-primed polymerization, reactions were supplemented by 0.5 mM ApG (TriLink BioTechnologies, San Diego, USA or IBA, Goettingen, Germany) or by 1 μM of capped-RNA (5΄-m7 GpppAAUCUAUAAUAG-3΄; TriLink BioTechnologies, San Diego, USA) if not indicated otherwise.

    Techniques: In Vitro, Concentration Assay, Standard Deviation

    Schematic work flow of the high-throughput compatible RNA synthesis assay. Influenza B polymerase (yellow sphere) activated by 5΄ RNA (nt 1–14) and bound to the 3΄ template RNA (labelled by FAM-Ex-5 at its 5΄ end) is incubated with nucleoside triphosphates (NTPs) and optionally a primer or generally a molecule X whose effect on RNA synthesis is to be monitored. The reactions are quenched by 4 M NaCl final which perturbs polymerase–RNA interactions but permits RNA–RNA interactions. Fluorescence polarization (FP) is recorded after equilibration (development) and directly reads out the ratio of full-length product RNA over labelled template RNA (see Supplementary Figure S3 ).

    Journal: Nucleic Acids Research

    Article Title: An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase

    doi: 10.1093/nar/gkx043

    Figure Lengend Snippet: Schematic work flow of the high-throughput compatible RNA synthesis assay. Influenza B polymerase (yellow sphere) activated by 5΄ RNA (nt 1–14) and bound to the 3΄ template RNA (labelled by FAM-Ex-5 at its 5΄ end) is incubated with nucleoside triphosphates (NTPs) and optionally a primer or generally a molecule X whose effect on RNA synthesis is to be monitored. The reactions are quenched by 4 M NaCl final which perturbs polymerase–RNA interactions but permits RNA–RNA interactions. Fluorescence polarization (FP) is recorded after equilibration (development) and directly reads out the ratio of full-length product RNA over labelled template RNA (see Supplementary Figure S3 ).

    Article Snippet: To monitor RNA-primed polymerization, reactions were supplemented by 0.5 mM ApG (TriLink BioTechnologies, San Diego, USA or IBA, Goettingen, Germany) or by 1 μM of capped-RNA (5΄-m7 GpppAAUCUAUAAUAG-3΄; TriLink BioTechnologies, San Diego, USA) if not indicated otherwise.

    Techniques: Flow Cytometry, High Throughput Screening Assay, Incubation, Fluorescence

    Structure of influenza B polymerase co-crystallized with the vRNA promoter an d capped primer. ( A ) Schematic of RNAs used for crystallization of the putative transcription initiation state. Nucleotides 4–13 of the 13-mer capped RNA primer are in italics as they are not seen in the structure. ( B ) Overall view of the crystal structure with influenza B polymerase in ribbon representation colored according to domains as indicated. The RNA in space-filling representation with the capped primer (blue), 5΄ end (violet) and 3΄ end (yellow), as in (A). ( C ) Trajectory of the 3΄ end (template) as observed in the current structure (yellow) and in the previous FluB2 structure (orange) ( 8 ). ( D ) As (C) but in the context of the polymerase (surface and ribbon representation) showing the template passing through the entry tunnel into the active site cavity. ( E ) Overall view of the promoter bound to the PB1 subunit (cyan ribbons) showing the priming loop (purple) and active site motif C (red). The unobserved 3΄ terminal base (U1, red) is modeled. Based on superposition with the Norwalk virus polymerase primer-template/substrate (CTP) complex (PDB: 3BSO, ( 30 )) the initial ApG of the product (orange), the incoming CTP (black) and the two catalytic divalent ions (green spheres) are modelled. ( F ) As (E) but only showing the RNA.

    Journal: Nucleic Acids Research

    Article Title: An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase

    doi: 10.1093/nar/gkx043

    Figure Lengend Snippet: Structure of influenza B polymerase co-crystallized with the vRNA promoter an d capped primer. ( A ) Schematic of RNAs used for crystallization of the putative transcription initiation state. Nucleotides 4–13 of the 13-mer capped RNA primer are in italics as they are not seen in the structure. ( B ) Overall view of the crystal structure with influenza B polymerase in ribbon representation colored according to domains as indicated. The RNA in space-filling representation with the capped primer (blue), 5΄ end (violet) and 3΄ end (yellow), as in (A). ( C ) Trajectory of the 3΄ end (template) as observed in the current structure (yellow) and in the previous FluB2 structure (orange) ( 8 ). ( D ) As (C) but in the context of the polymerase (surface and ribbon representation) showing the template passing through the entry tunnel into the active site cavity. ( E ) Overall view of the promoter bound to the PB1 subunit (cyan ribbons) showing the priming loop (purple) and active site motif C (red). The unobserved 3΄ terminal base (U1, red) is modeled. Based on superposition with the Norwalk virus polymerase primer-template/substrate (CTP) complex (PDB: 3BSO, ( 30 )) the initial ApG of the product (orange), the incoming CTP (black) and the two catalytic divalent ions (green spheres) are modelled. ( F ) As (E) but only showing the RNA.

    Article Snippet: To monitor RNA-primed polymerization, reactions were supplemented by 0.5 mM ApG (TriLink BioTechnologies, San Diego, USA or IBA, Goettingen, Germany) or by 1 μM of capped-RNA (5΄-m7 GpppAAUCUAUAAUAG-3΄; TriLink BioTechnologies, San Diego, USA) if not indicated otherwise.

    Techniques: Crystallization Assay

    Terminal (vPol) versus internal (cPol) initiation of RNA synthesis. ( A ) The vRNA promoter (vPol; black; 3 independent rate-NTP-characteristics, circles, upward and downward triangles) or the cRNA promoter (cPol; pink; 2 independent rate-NTP-characteristics, upward and downward triangles) direct the initiation of RNA synthesis by influenza B polymerase with different enzymatic parameters (Table 1 ). The dependency of RNA synthesis (observed rate constants) on the substrate-concentration (NTP) was fitted according to a simple substrate-inhibition model proposed by HALDANE ( 19 ) (see Materials and Methods). The promoter RNA sequences responsible for the altered k cat , K M (NTPs) and K M (primer) are shown in the lower panel. Both the v5΄ and the c5΄ comprise a proximal hook structure (nt 1–10; blue colouring) followed by the distal 5΄-3΄ base-pairing region (nts 11–13/14; green colouring). The v3΄ and c3΄ template RNAs each direct the incorporation of NTPs based on complementarity. ( B ) Deletion of the ultimate U1 from the template RNA impairs initiation of RNA synthesis from the vRNA promoter but not cRNA promoter, indicating terminal and internal initiation mechanisms, respectively. Shown are five independent progress curves each for initiation at the vRNA and cRNA promoter (black and pink) and the corresponding U1 deletions, respectively (purple and orange). ( C ) vPol-like kinetics (black) were recovered by a cPol with either (i) A3 of c3΄ substituted by G3, as found in vRNA (c3΄A3G; green); or (ii) the distal cRNA promoter involving the 5΄-3΄ base-pairing region being swapped for the corresponding region of the vRNA promoter (5΄ nt 11–18, 3΄ nt 10–18; c3΄ v bp , purple); or (iii) the proximal cRNA promoter swapped for the corresponding region of the vRNA promoter (5΄ nt 1–10, 3΄ nt 1–9; v3΄ c bp , orange) (see Supplementary Figure S12 ). Only the c3΄ template RNA in combination with the cRNA promoter's 5΄-3΄ base-pairing region resulted in cPol-like slow progress curves indicating internal initiation (pink). The bi-phasic kinetics of RNA synthesis by some RNA combinations was attributed to strengthened RNA–RNA interactions which kinetically competed with polymerase for single-stranded template RNA (Figure 1 , Supplementary Figure S13 ). In each case, five independent kinetics are shown.

    Journal: Nucleic Acids Research

    Article Title: An in vitro fluorescence based study of initiation of RNA synthesis by influenza B polymerase

    doi: 10.1093/nar/gkx043

    Figure Lengend Snippet: Terminal (vPol) versus internal (cPol) initiation of RNA synthesis. ( A ) The vRNA promoter (vPol; black; 3 independent rate-NTP-characteristics, circles, upward and downward triangles) or the cRNA promoter (cPol; pink; 2 independent rate-NTP-characteristics, upward and downward triangles) direct the initiation of RNA synthesis by influenza B polymerase with different enzymatic parameters (Table 1 ). The dependency of RNA synthesis (observed rate constants) on the substrate-concentration (NTP) was fitted according to a simple substrate-inhibition model proposed by HALDANE ( 19 ) (see Materials and Methods). The promoter RNA sequences responsible for the altered k cat , K M (NTPs) and K M (primer) are shown in the lower panel. Both the v5΄ and the c5΄ comprise a proximal hook structure (nt 1–10; blue colouring) followed by the distal 5΄-3΄ base-pairing region (nts 11–13/14; green colouring). The v3΄ and c3΄ template RNAs each direct the incorporation of NTPs based on complementarity. ( B ) Deletion of the ultimate U1 from the template RNA impairs initiation of RNA synthesis from the vRNA promoter but not cRNA promoter, indicating terminal and internal initiation mechanisms, respectively. Shown are five independent progress curves each for initiation at the vRNA and cRNA promoter (black and pink) and the corresponding U1 deletions, respectively (purple and orange). ( C ) vPol-like kinetics (black) were recovered by a cPol with either (i) A3 of c3΄ substituted by G3, as found in vRNA (c3΄A3G; green); or (ii) the distal cRNA promoter involving the 5΄-3΄ base-pairing region being swapped for the corresponding region of the vRNA promoter (5΄ nt 11–18, 3΄ nt 10–18; c3΄ v bp , purple); or (iii) the proximal cRNA promoter swapped for the corresponding region of the vRNA promoter (5΄ nt 1–10, 3΄ nt 1–9; v3΄ c bp , orange) (see Supplementary Figure S12 ). Only the c3΄ template RNA in combination with the cRNA promoter's 5΄-3΄ base-pairing region resulted in cPol-like slow progress curves indicating internal initiation (pink). The bi-phasic kinetics of RNA synthesis by some RNA combinations was attributed to strengthened RNA–RNA interactions which kinetically competed with polymerase for single-stranded template RNA (Figure 1 , Supplementary Figure S13 ). In each case, five independent kinetics are shown.

    Article Snippet: To monitor RNA-primed polymerization, reactions were supplemented by 0.5 mM ApG (TriLink BioTechnologies, San Diego, USA or IBA, Goettingen, Germany) or by 1 μM of capped-RNA (5΄-m7 GpppAAUCUAUAAUAG-3΄; TriLink BioTechnologies, San Diego, USA) if not indicated otherwise.

    Techniques: Concentration Assay, Inhibition