rna loading buffer Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86
    Thermo Fisher rna loading buffer
    Length-dependence of poly(A)- and SRE-effects assayed in a series of <t>RNAs</t> based on nanoluciferase. ( A ) Scheme of the RNAs used. All contained an m7G cap and the nLuc ORF. They had poly(A) tails and/or SREs at their 3′ end as shown. All SRE-containing RNAs were synthesized in an SRE + and an SRE − version as indicated. The two RNAs at the bottom were extended by the ORF encoding MBP (orange line) inserted between the nLuc stop codon and the SREs. “N” indicates the number of independent <t>RNA</t> preparations assayed. Each of the three preparations of each RNA was tested in two to four independent batches of Drosophila embryo extract, resulting in nine independent experiments per RNA (“n”). ( B ) Summary of translation experiments carried out with the RNAs depicted above . Poly(A) stimulation, shown on the left , was calculated from the comparison of equivalent SRE − RNAs carrying a poly(A) stretch or not ( P ≤ 3.8 × 10 −5 ). SRE-dependent repression, shown on the right , was calculated from the comparison of SRE + and SRE − RNAs that also carried a poly(A) tail ( P ≤ 5.8 × 10 −4 ). Both stimulation and repression are shown with a correction for translational efficiency in RRL (red bars) and without correction (blue bars). Correction factors in individual experiments were typically between 1 and 1.5 and up to 3 in a few cases. Error bars indicate the standard deviation based on the numbers of independent experiments reported in A . The statistical significance of the pairwise comparisons indicated by brackets is indicated by (*) ( P
    Rna Loading Buffer, supplied by Thermo Fisher, 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/rna loading buffer/product/Thermo Fisher
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rna loading buffer - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    97
    Millipore rna loading buffer
    Length-dependence of poly(A)- and SRE-effects assayed in a series of <t>RNAs</t> based on nanoluciferase. ( A ) Scheme of the RNAs used. All contained an m7G cap and the nLuc ORF. They had poly(A) tails and/or SREs at their 3′ end as shown. All SRE-containing RNAs were synthesized in an SRE + and an SRE − version as indicated. The two RNAs at the bottom were extended by the ORF encoding MBP (orange line) inserted between the nLuc stop codon and the SREs. “N” indicates the number of independent <t>RNA</t> preparations assayed. Each of the three preparations of each RNA was tested in two to four independent batches of Drosophila embryo extract, resulting in nine independent experiments per RNA (“n”). ( B ) Summary of translation experiments carried out with the RNAs depicted above . Poly(A) stimulation, shown on the left , was calculated from the comparison of equivalent SRE − RNAs carrying a poly(A) stretch or not ( P ≤ 3.8 × 10 −5 ). SRE-dependent repression, shown on the right , was calculated from the comparison of SRE + and SRE − RNAs that also carried a poly(A) tail ( P ≤ 5.8 × 10 −4 ). Both stimulation and repression are shown with a correction for translational efficiency in RRL (red bars) and without correction (blue bars). Correction factors in individual experiments were typically between 1 and 1.5 and up to 3 in a few cases. Error bars indicate the standard deviation based on the numbers of independent experiments reported in A . The statistical significance of the pairwise comparisons indicated by brackets is indicated by (*) ( P
    Rna Loading Buffer, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rna loading buffer/product/Millipore
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rna loading buffer - by Bioz Stars, 2021-06
    97/100 stars
      Buy from Supplier

    97
    Thermo Fisher 1x loading buffer
    Length-dependence of poly(A)- and SRE-effects assayed in a series of <t>RNAs</t> based on nanoluciferase. ( A ) Scheme of the RNAs used. All contained an m7G cap and the nLuc ORF. They had poly(A) tails and/or SREs at their 3′ end as shown. All SRE-containing RNAs were synthesized in an SRE + and an SRE − version as indicated. The two RNAs at the bottom were extended by the ORF encoding MBP (orange line) inserted between the nLuc stop codon and the SREs. “N” indicates the number of independent <t>RNA</t> preparations assayed. Each of the three preparations of each RNA was tested in two to four independent batches of Drosophila embryo extract, resulting in nine independent experiments per RNA (“n”). ( B ) Summary of translation experiments carried out with the RNAs depicted above . Poly(A) stimulation, shown on the left , was calculated from the comparison of equivalent SRE − RNAs carrying a poly(A) stretch or not ( P ≤ 3.8 × 10 −5 ). SRE-dependent repression, shown on the right , was calculated from the comparison of SRE + and SRE − RNAs that also carried a poly(A) tail ( P ≤ 5.8 × 10 −4 ). Both stimulation and repression are shown with a correction for translational efficiency in RRL (red bars) and without correction (blue bars). Correction factors in individual experiments were typically between 1 and 1.5 and up to 3 in a few cases. Error bars indicate the standard deviation based on the numbers of independent experiments reported in A . The statistical significance of the pairwise comparisons indicated by brackets is indicated by (*) ( P
    1x Loading Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/1x loading buffer/product/Thermo Fisher
    Average 97 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    1x loading buffer - by Bioz Stars, 2021-06
    97/100 stars
      Buy from Supplier

    99
    Thermo Fisher buffer ii
    Length-dependence of poly(A)- and SRE-effects assayed in a series of <t>RNAs</t> based on nanoluciferase. ( A ) Scheme of the RNAs used. All contained an m7G cap and the nLuc ORF. They had poly(A) tails and/or SREs at their 3′ end as shown. All SRE-containing RNAs were synthesized in an SRE + and an SRE − version as indicated. The two RNAs at the bottom were extended by the ORF encoding MBP (orange line) inserted between the nLuc stop codon and the SREs. “N” indicates the number of independent <t>RNA</t> preparations assayed. Each of the three preparations of each RNA was tested in two to four independent batches of Drosophila embryo extract, resulting in nine independent experiments per RNA (“n”). ( B ) Summary of translation experiments carried out with the RNAs depicted above . Poly(A) stimulation, shown on the left , was calculated from the comparison of equivalent SRE − RNAs carrying a poly(A) stretch or not ( P ≤ 3.8 × 10 −5 ). SRE-dependent repression, shown on the right , was calculated from the comparison of SRE + and SRE − RNAs that also carried a poly(A) tail ( P ≤ 5.8 × 10 −4 ). Both stimulation and repression are shown with a correction for translational efficiency in RRL (red bars) and without correction (blue bars). Correction factors in individual experiments were typically between 1 and 1.5 and up to 3 in a few cases. Error bars indicate the standard deviation based on the numbers of independent experiments reported in A . The statistical significance of the pairwise comparisons indicated by brackets is indicated by (*) ( P
    Buffer Ii, 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/buffer ii/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    buffer ii - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

    Image Search Results


    Length-dependence of poly(A)- and SRE-effects assayed in a series of RNAs based on nanoluciferase. ( A ) Scheme of the RNAs used. All contained an m7G cap and the nLuc ORF. They had poly(A) tails and/or SREs at their 3′ end as shown. All SRE-containing RNAs were synthesized in an SRE + and an SRE − version as indicated. The two RNAs at the bottom were extended by the ORF encoding MBP (orange line) inserted between the nLuc stop codon and the SREs. “N” indicates the number of independent RNA preparations assayed. Each of the three preparations of each RNA was tested in two to four independent batches of Drosophila embryo extract, resulting in nine independent experiments per RNA (“n”). ( B ) Summary of translation experiments carried out with the RNAs depicted above . Poly(A) stimulation, shown on the left , was calculated from the comparison of equivalent SRE − RNAs carrying a poly(A) stretch or not ( P ≤ 3.8 × 10 −5 ). SRE-dependent repression, shown on the right , was calculated from the comparison of SRE + and SRE − RNAs that also carried a poly(A) tail ( P ≤ 5.8 × 10 −4 ). Both stimulation and repression are shown with a correction for translational efficiency in RRL (red bars) and without correction (blue bars). Correction factors in individual experiments were typically between 1 and 1.5 and up to 3 in a few cases. Error bars indicate the standard deviation based on the numbers of independent experiments reported in A . The statistical significance of the pairwise comparisons indicated by brackets is indicated by (*) ( P

    Journal: RNA

    Article Title: Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone

    doi: 10.1261/rna.073759.119

    Figure Lengend Snippet: Length-dependence of poly(A)- and SRE-effects assayed in a series of RNAs based on nanoluciferase. ( A ) Scheme of the RNAs used. All contained an m7G cap and the nLuc ORF. They had poly(A) tails and/or SREs at their 3′ end as shown. All SRE-containing RNAs were synthesized in an SRE + and an SRE − version as indicated. The two RNAs at the bottom were extended by the ORF encoding MBP (orange line) inserted between the nLuc stop codon and the SREs. “N” indicates the number of independent RNA preparations assayed. Each of the three preparations of each RNA was tested in two to four independent batches of Drosophila embryo extract, resulting in nine independent experiments per RNA (“n”). ( B ) Summary of translation experiments carried out with the RNAs depicted above . Poly(A) stimulation, shown on the left , was calculated from the comparison of equivalent SRE − RNAs carrying a poly(A) stretch or not ( P ≤ 3.8 × 10 −5 ). SRE-dependent repression, shown on the right , was calculated from the comparison of SRE + and SRE − RNAs that also carried a poly(A) tail ( P ≤ 5.8 × 10 −4 ). Both stimulation and repression are shown with a correction for translational efficiency in RRL (red bars) and without correction (blue bars). Correction factors in individual experiments were typically between 1 and 1.5 and up to 3 in a few cases. Error bars indicate the standard deviation based on the numbers of independent experiments reported in A . The statistical significance of the pairwise comparisons indicated by brackets is indicated by (*) ( P

    Article Snippet: RNAs were ethanol-precipitated, dissolved in RNA-loading-buffer (Thermo Scientific), and resolved on denaturing agarose gels containing 1× MOPS buffer and 2.2 M formaldehyde ( ).

    Techniques: Synthesized, Standard Deviation

    Analysis of the SRE-dependent repressor complex assembled on chimeric RNAs. ( A ) Rapid sedimentation of repressed RNAs. Radiolabeled chimeric RNAs were incubated for 30 min in Drosophila embryo extract under preincubation conditions. Radiolabeled regulatory RNAs not fused to the nLuc reporter ( top left ). ( B ) Partial RNase resistance of repressed RNAs. Regulatory RNA by itself as well as flipped and forward chimeric constructs, all in the SRE + and SRE − versions, were incubated in Drosophila embryo extract under preincubation conditions, then a time-course of RNase digestion was performed as described in Materials and Methods. Time points were taken at 5 min intervals. A radiolabeled synthetic RNA oligonucleotide was added before RNA purification and served as a recovery control ( bottom part of the gel). White and black arrowheads indicate the nLuc and the regulatory RNA, respectively. Stabilization factors were calculated by comparison of the SRE + RNA and SRE − RNA half-lives and are listed at the bottom .

    Journal: RNA

    Article Title: Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone

    doi: 10.1261/rna.073759.119

    Figure Lengend Snippet: Analysis of the SRE-dependent repressor complex assembled on chimeric RNAs. ( A ) Rapid sedimentation of repressed RNAs. Radiolabeled chimeric RNAs were incubated for 30 min in Drosophila embryo extract under preincubation conditions. Radiolabeled regulatory RNAs not fused to the nLuc reporter ( top left ). ( B ) Partial RNase resistance of repressed RNAs. Regulatory RNA by itself as well as flipped and forward chimeric constructs, all in the SRE + and SRE − versions, were incubated in Drosophila embryo extract under preincubation conditions, then a time-course of RNase digestion was performed as described in Materials and Methods. Time points were taken at 5 min intervals. A radiolabeled synthetic RNA oligonucleotide was added before RNA purification and served as a recovery control ( bottom part of the gel). White and black arrowheads indicate the nLuc and the regulatory RNA, respectively. Stabilization factors were calculated by comparison of the SRE + RNA and SRE − RNA half-lives and are listed at the bottom .

    Article Snippet: RNAs were ethanol-precipitated, dissolved in RNA-loading-buffer (Thermo Scientific), and resolved on denaturing agarose gels containing 1× MOPS buffer and 2.2 M formaldehyde ( ).

    Techniques: Sedimentation, Incubation, Construct, Purification

    Assembly of chimeric RNA constructs. ( A ) Outline of the procedure. Regulatory RNAs contained a 5′ biotin, two SREs (SRE + = WT or SRE − , that is, with an inactivating point mutation in each SRE) and a poly(A) sequence, which was protected against deadenylation by an N 40 sequence at the 3′ end. Alternatively, regulatory RNA was used that carried a poly(A) sequence but no SREs (not shown). The regulatory RNA was hybridized to immobilized oligo(dT), then divalent streptavidin and the capped and 3′-biotinylated nLuc reporter RNA were added in a stepwise manner. The chimeric construct was eluted in low-salt buffer. The scheme shows the assembly of forward constructs. Flipped constructs were assembled in the same manner, but 3′-biotinylated regulatory RNAs were used. For a full description of the procedure, see Materials and Methods. ( B ) Assembly of chimeric RNAs assayed by native gel electrophoresis. Chimeras [forward-SRE ± p(A)] were assembled as described in panel A and Materials and Methods. The figure shows the analysis of input and product RNAs by electrophoresis through a 5% nondenaturing polyacrylamide gel. Lanes 1 – 3 display the input RNAs as indicated. Lanes 4 – 6 demonstrate the retardation of input RNAs upon mixing with divalent streptavidin (STV). Two retarded bands in one lane are presumably due to binding of one or two RNAs to one molecule of streptavidin. Lanes 7 , 8 show the flow-through of the oligo(dT) matrix after loading with regulatory RNAs; binding was essentially complete. Lanes 9 and 10 show the flow-through of excess nLuc RNA in the last assembly step. “nLuc + ”′ and “nLuc − ” refer to the nLuc RNAs from the assembly reactions with SRE + and SRE − regulatory RNAs, respectively. Lanes 11 and 12 show the eluate in formamide-containing loading buffer without heat denaturation. The main band (arrowhead) is a novel species that we interpret as the desired chimeric RNA. Upon heat denaturation (lanes 13 , 14 ), the input RNAs are restored. Residual amounts of input RNAs can be detected: Based on a comparison to other lanes in this gel, numbered bands are (1) a complex between one regulatory RNA and streptavidin, (2) bare nLuc RNA, and (3) a complex between two regulatory RNAs and streptavidin. The broken line indicates the upper end of the gel.

    Journal: RNA

    Article Title: Establishment of 5′–3′ interactions in mRNA independent of a continuous ribose-phosphate backbone

    doi: 10.1261/rna.073759.119

    Figure Lengend Snippet: Assembly of chimeric RNA constructs. ( A ) Outline of the procedure. Regulatory RNAs contained a 5′ biotin, two SREs (SRE + = WT or SRE − , that is, with an inactivating point mutation in each SRE) and a poly(A) sequence, which was protected against deadenylation by an N 40 sequence at the 3′ end. Alternatively, regulatory RNA was used that carried a poly(A) sequence but no SREs (not shown). The regulatory RNA was hybridized to immobilized oligo(dT), then divalent streptavidin and the capped and 3′-biotinylated nLuc reporter RNA were added in a stepwise manner. The chimeric construct was eluted in low-salt buffer. The scheme shows the assembly of forward constructs. Flipped constructs were assembled in the same manner, but 3′-biotinylated regulatory RNAs were used. For a full description of the procedure, see Materials and Methods. ( B ) Assembly of chimeric RNAs assayed by native gel electrophoresis. Chimeras [forward-SRE ± p(A)] were assembled as described in panel A and Materials and Methods. The figure shows the analysis of input and product RNAs by electrophoresis through a 5% nondenaturing polyacrylamide gel. Lanes 1 – 3 display the input RNAs as indicated. Lanes 4 – 6 demonstrate the retardation of input RNAs upon mixing with divalent streptavidin (STV). Two retarded bands in one lane are presumably due to binding of one or two RNAs to one molecule of streptavidin. Lanes 7 , 8 show the flow-through of the oligo(dT) matrix after loading with regulatory RNAs; binding was essentially complete. Lanes 9 and 10 show the flow-through of excess nLuc RNA in the last assembly step. “nLuc + ”′ and “nLuc − ” refer to the nLuc RNAs from the assembly reactions with SRE + and SRE − regulatory RNAs, respectively. Lanes 11 and 12 show the eluate in formamide-containing loading buffer without heat denaturation. The main band (arrowhead) is a novel species that we interpret as the desired chimeric RNA. Upon heat denaturation (lanes 13 , 14 ), the input RNAs are restored. Residual amounts of input RNAs can be detected: Based on a comparison to other lanes in this gel, numbered bands are (1) a complex between one regulatory RNA and streptavidin, (2) bare nLuc RNA, and (3) a complex between two regulatory RNAs and streptavidin. The broken line indicates the upper end of the gel.

    Article Snippet: RNAs were ethanol-precipitated, dissolved in RNA-loading-buffer (Thermo Scientific), and resolved on denaturing agarose gels containing 1× MOPS buffer and 2.2 M formaldehyde ( ).

    Techniques: Construct, Mutagenesis, Sequencing, Nucleic Acid Electrophoresis, Electrophoresis, Binding Assay