in vitro run-off transcription Search Results


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  • 99
    New England Biolabs nsii
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    Thermo Fisher run off transcription in vitro
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    Roche run off in vitro transcription
    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
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    Promega in vitro run off transcription reaction
    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
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    Roche t7 rnap
    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
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    Promega riboprobe
    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
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    Thermo Fisher megashortscript transcription kit
    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
    Megashortscript Transcription Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 240 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher mmessage mmachine sp6 transcription kit
    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
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    Thermo Fisher t3 mmessage mmachine kit
    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
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    Outline of the first subtractive step. Synthetic RNAs were produced by <t>run-off</t> <t>in</t> <t>vitro</t> <t>transcription</t> from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.
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    New England Biolabs t7 rna polymerase
    Opposite co-transcriptional folding paths of a pair of RNA switches with ‘direct’ and ‘reverse’ sequences (i.e. 5′-ABCD-3′ versus 5′-DCBA-3′). Structures 1D and 1R (respectively, 2D and 2R) of the direct and reverse switches are energetically equivalent because of helix symmetries; dashed lines indicate mirror symmetry of Pa, Pb, Pc and Pd which are therefore conserved under sequence reversal relating direct and reverse switches. Despite these strong similarities between D and R structures at equilibrium, direct and reverse switches display ‘opposite’ co-transcriptional folding paths (direct switch into structure 1D and reverse switch into structure 2R) guided through a helix encoded persistence (left) or exchange (right) during in vitro transcription using <t>T7</t> RNA polymerase (see Materials and Methods).
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    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Thermo Fisher t7 rna polymerase promoters
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Promega t7 rna polymerase kit
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Thermo Fisher t3 megascript kit
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Millipore dig rna labeling kit
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Thermo Fisher t7 mmessage mmachine kit
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Thermo Fisher megascript sp6 kit
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Promega ribomax kit
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Thermo Fisher t7 megashortscript
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    New England Biolabs scai hf
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Millipore dna templates
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Thermo Fisher transcriptaidtm t7 high yield transcription kit
    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. <t>T7</t> RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.
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    Image Search Results


    Outline of the first subtractive step. Synthetic RNAs were produced by run-off in vitro transcription from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.

    Journal: Nucleic Acids Research

    Article Title: A two-step strategy for constructing specifically self-subtracted cDNA libraries

    doi:

    Figure Lengend Snippet: Outline of the first subtractive step. Synthetic RNAs were produced by run-off in vitro transcription from a 3′-end-specific cDNA library (HM3). The resulting RNA molecules are nearly uniform in length (400–600 nt) and their composition reflects the relative abundance of each cDNA in the starting library. Super-prevalent RNAs are shown as black wavy lines. We designed DNA oligonucleotides complementary to the 3′-UTR of the super-prevalent mRNAs, then RNAs and DNA oligonucleotides were incubated in the presence of RNase H, which specifically degrades the RNA moiety in a DNA:RNA hybrid. The undigested RNAs were purified from preparative gels, reverse transcribed and cloned in plasmid vectors.

    Article Snippet: Antisense, biotinylated RNA molecules were obtained by run-off in vitro transcription in the presence of biotin-16-UTP (Roche).

    Techniques: Produced, In Vitro, cDNA Library Assay, Incubation, Purification, Clone Assay, Plasmid Preparation

    Opposite co-transcriptional folding paths of a pair of RNA switches with ‘direct’ and ‘reverse’ sequences (i.e. 5′-ABCD-3′ versus 5′-DCBA-3′). Structures 1D and 1R (respectively, 2D and 2R) of the direct and reverse switches are energetically equivalent because of helix symmetries; dashed lines indicate mirror symmetry of Pa, Pb, Pc and Pd which are therefore conserved under sequence reversal relating direct and reverse switches. Despite these strong similarities between D and R structures at equilibrium, direct and reverse switches display ‘opposite’ co-transcriptional folding paths (direct switch into structure 1D and reverse switch into structure 2R) guided through a helix encoded persistence (left) or exchange (right) during in vitro transcription using T7 RNA polymerase (see Materials and Methods).

    Journal: Nucleic Acids Research

    Article Title: Encoding folding paths of RNA switches

    doi: 10.1093/nar/gkl1036

    Figure Lengend Snippet: Opposite co-transcriptional folding paths of a pair of RNA switches with ‘direct’ and ‘reverse’ sequences (i.e. 5′-ABCD-3′ versus 5′-DCBA-3′). Structures 1D and 1R (respectively, 2D and 2R) of the direct and reverse switches are energetically equivalent because of helix symmetries; dashed lines indicate mirror symmetry of Pa, Pb, Pc and Pd which are therefore conserved under sequence reversal relating direct and reverse switches. Despite these strong similarities between D and R structures at equilibrium, direct and reverse switches display ‘opposite’ co-transcriptional folding paths (direct switch into structure 1D and reverse switch into structure 2R) guided through a helix encoded persistence (left) or exchange (right) during in vitro transcription using T7 RNA polymerase (see Materials and Methods).

    Article Snippet: Run off transcription was performed in vitro using T7 RNA polymerase (New England Biolabs) for up to 4–5 h at 37 or 25°C.

    Techniques: Sequencing, In Vitro

    Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. T7 RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.

    Journal: Nucleic Acids Research

    Article Title: Monitoring mis-acylated tRNA suppression efficiency in mammalian cells via EGFP fluorescence recovery

    doi:

    Figure Lengend Snippet: Scheme for the in vitro synthesis of the suppressor tRNA from a synthetic DNA template by T7 run-off transcription. Two synthetic complementary oligonucleotides encoding the E.coli -derived suppressor tRNA Ala CUA were annealed and cloned into the pCR4 vector, leading to pEcoliAlaCUA. T7 RNA polymerase run-off transcription was performed on the purified Eco RI and Bsa I fragment giving rise to a 76mer suppressor tRNA.

    Article Snippet: After agarose gel purification this DNA fragment was used for the run-off transcription with T7 RNA polymerase following the protocol of the MegaScript kit (Ambion).

    Techniques: In Vitro, Derivative Assay, Clone Assay, Plasmid Preparation, Purification