rna  (New England Biolabs)


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  • 93
    Name:
    Vaccinia Capping System
    Description:
    Vaccinia Capping System 400 units
    Catalog Number:
    m2080s
    Price:
    140
    Size:
    400 units
    Category:
    Capping Reagents for DNA RNA Synthesis
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    Structured Review

    New England Biolabs rna
    Vaccinia Capping System
    Vaccinia Capping System 400 units
    https://www.bioz.com/result/rna/product/New England Biolabs
    Average 93 stars, based on 130 article reviews
    Price from $9.99 to $1999.99
    rna - by Bioz Stars, 2020-07
    93/100 stars

    Images

    1) Product Images from "Global repositioning of transcription start sites in a plant-fermenting bacterium"

    Article Title: Global repositioning of transcription start sites in a plant-fermenting bacterium

    Journal: Nature Communications

    doi: 10.1038/ncomms13783

    Overview of the Capp-Switch sequencing approach. Capp-Switch includes ( a – c ) capture of 5′ mRNA fragments and ( d – f ) cDNA synthesis and sequencing. ( a ) The mRNA 5′ triphosphate is capped with biotin-GTP by VCE. ( b ) RNA is fragmented and ( c ) the capped 5′ mRNA fragments are captured on streptavidin magnetic beads and separated from other RNA. ( d ) The 5′ mRNA fragments are reverse transcribed to single-stranded cDNA using MMLV reverse transcriptase. An oligonucleotide hybridizes to the 3′ overhang and the complementary sequence is synthesized by the MMLV template-switching activity. ( e ) Double-stranded cDNA is synthesized using primers that hybridize to the single-stranded cDNA termini. ( f ) The cDNA is sequenced on a high-throughput platform.
    Figure Legend Snippet: Overview of the Capp-Switch sequencing approach. Capp-Switch includes ( a – c ) capture of 5′ mRNA fragments and ( d – f ) cDNA synthesis and sequencing. ( a ) The mRNA 5′ triphosphate is capped with biotin-GTP by VCE. ( b ) RNA is fragmented and ( c ) the capped 5′ mRNA fragments are captured on streptavidin magnetic beads and separated from other RNA. ( d ) The 5′ mRNA fragments are reverse transcribed to single-stranded cDNA using MMLV reverse transcriptase. An oligonucleotide hybridizes to the 3′ overhang and the complementary sequence is synthesized by the MMLV template-switching activity. ( e ) Double-stranded cDNA is synthesized using primers that hybridize to the single-stranded cDNA termini. ( f ) The cDNA is sequenced on a high-throughput platform.

    Techniques Used: Sequencing, Magnetic Beads, Synthesized, Activity Assay, High Throughput Screening Assay

    2) Product Images from "Global repositioning of transcription start sites in a plant-fermenting bacterium"

    Article Title: Global repositioning of transcription start sites in a plant-fermenting bacterium

    Journal: Nature Communications

    doi: 10.1038/ncomms13783

    Overview of the Capp-Switch sequencing approach. Capp-Switch includes ( a – c ) capture of 5′ mRNA fragments and ( d – f ) cDNA synthesis and sequencing. ( a ) The mRNA 5′ triphosphate is capped with biotin-GTP by VCE. ( b ) RNA is fragmented and ( c ) the capped 5′ mRNA fragments are captured on streptavidin magnetic beads and separated from other RNA. ( d ) The 5′ mRNA fragments are reverse transcribed to single-stranded cDNA using MMLV reverse transcriptase. An oligonucleotide hybridizes to the 3′ overhang and the complementary sequence is synthesized by the MMLV template-switching activity. ( e ) Double-stranded cDNA is synthesized using primers that hybridize to the single-stranded cDNA termini. ( f ) The cDNA is sequenced on a high-throughput platform.
    Figure Legend Snippet: Overview of the Capp-Switch sequencing approach. Capp-Switch includes ( a – c ) capture of 5′ mRNA fragments and ( d – f ) cDNA synthesis and sequencing. ( a ) The mRNA 5′ triphosphate is capped with biotin-GTP by VCE. ( b ) RNA is fragmented and ( c ) the capped 5′ mRNA fragments are captured on streptavidin magnetic beads and separated from other RNA. ( d ) The 5′ mRNA fragments are reverse transcribed to single-stranded cDNA using MMLV reverse transcriptase. An oligonucleotide hybridizes to the 3′ overhang and the complementary sequence is synthesized by the MMLV template-switching activity. ( e ) Double-stranded cDNA is synthesized using primers that hybridize to the single-stranded cDNA termini. ( f ) The cDNA is sequenced on a high-throughput platform.

    Techniques Used: Sequencing, Magnetic Beads, Synthesized, Activity Assay, High Throughput Screening Assay

    Related Articles

    Transfection:

    Article Title: La-related protein 1 (LARP1) repression of TOP mRNA translation is mediated through its cap-binding domain and controlled by an adjacent regulatory region
    Article Snippet: .. Materials Reagents were obtained from the following sources: antibodies for S6K, phospho-T389-S6K, eIF2α, phospho-Ser51-eIF2α, Raptor, mTOR, 4EBP1, LARP1, NCBP1, eIF4E, eIF4G and PABP from Cell Signaling Technology; primary antibodies for eIF3b and HRP-labeled secondary antibodies from Santa Cruz Biotechnology; IRDye secondary antibodies from LI-COR; Dulbecco’s modified Eagle’s medium (DMEM) from Life Technologies; heat-inactivated Fetal Bovine Serum (IFS) and 7mGDP from Sigma Aldrich; DNase I, T4 DNA ligase 1, T4 RNA Ligase I, T7 RNA polymerase, polynucleotide kinase, Protoscript II reverse transcriptase, Vaccinia Capping System, Oligo d(T)25 Magnetic beads and streptavidin-coated magnetic beads from New England Biolabs; iTaq Universal SYBR Green Supermix and Bradford Protein Assay from Bio-rad; RNeasy Plus Mini Kit from Qiagen; Dual Luciferase Assay from Promega; and X-tremeGENE 9 transfection reagent from Roche. .. Cell culture and preparation of cell extracts Cells were grown in high-glucose DMEM supplemented with 10% (v/v) heat-inactivated fetal bovine serum and penicillin-streptomycin.

    Luciferase:

    Article Title: La-related protein 1 (LARP1) repression of TOP mRNA translation is mediated through its cap-binding domain and controlled by an adjacent regulatory region
    Article Snippet: .. Materials Reagents were obtained from the following sources: antibodies for S6K, phospho-T389-S6K, eIF2α, phospho-Ser51-eIF2α, Raptor, mTOR, 4EBP1, LARP1, NCBP1, eIF4E, eIF4G and PABP from Cell Signaling Technology; primary antibodies for eIF3b and HRP-labeled secondary antibodies from Santa Cruz Biotechnology; IRDye secondary antibodies from LI-COR; Dulbecco’s modified Eagle’s medium (DMEM) from Life Technologies; heat-inactivated Fetal Bovine Serum (IFS) and 7mGDP from Sigma Aldrich; DNase I, T4 DNA ligase 1, T4 RNA Ligase I, T7 RNA polymerase, polynucleotide kinase, Protoscript II reverse transcriptase, Vaccinia Capping System, Oligo d(T)25 Magnetic beads and streptavidin-coated magnetic beads from New England Biolabs; iTaq Universal SYBR Green Supermix and Bradford Protein Assay from Bio-rad; RNeasy Plus Mini Kit from Qiagen; Dual Luciferase Assay from Promega; and X-tremeGENE 9 transfection reagent from Roche. .. Cell culture and preparation of cell extracts Cells were grown in high-glucose DMEM supplemented with 10% (v/v) heat-inactivated fetal bovine serum and penicillin-streptomycin.

    Magnetic Beads:

    Article Title: La-related protein 1 (LARP1) repression of TOP mRNA translation is mediated through its cap-binding domain and controlled by an adjacent regulatory region
    Article Snippet: .. Materials Reagents were obtained from the following sources: antibodies for S6K, phospho-T389-S6K, eIF2α, phospho-Ser51-eIF2α, Raptor, mTOR, 4EBP1, LARP1, NCBP1, eIF4E, eIF4G and PABP from Cell Signaling Technology; primary antibodies for eIF3b and HRP-labeled secondary antibodies from Santa Cruz Biotechnology; IRDye secondary antibodies from LI-COR; Dulbecco’s modified Eagle’s medium (DMEM) from Life Technologies; heat-inactivated Fetal Bovine Serum (IFS) and 7mGDP from Sigma Aldrich; DNase I, T4 DNA ligase 1, T4 RNA Ligase I, T7 RNA polymerase, polynucleotide kinase, Protoscript II reverse transcriptase, Vaccinia Capping System, Oligo d(T)25 Magnetic beads and streptavidin-coated magnetic beads from New England Biolabs; iTaq Universal SYBR Green Supermix and Bradford Protein Assay from Bio-rad; RNeasy Plus Mini Kit from Qiagen; Dual Luciferase Assay from Promega; and X-tremeGENE 9 transfection reagent from Roche. .. Cell culture and preparation of cell extracts Cells were grown in high-glucose DMEM supplemented with 10% (v/v) heat-inactivated fetal bovine serum and penicillin-streptomycin.

    Isolation:

    Article Title: Elimination of Ribosome Inactivating Factors Improves the Efficiency of Bacillus subtilis and Saccharomyces cerevisiae Cell-Free Translation Systems
    Article Snippet: .. A typical 20 μL reaction containing 1 pmole DNA was incubated for 2 h at 37°C prior to mRNA isolation with MEGAclearTM Kit (Ambion), followed by capping by the Vaccinia Capping System (NEB) and re-purification of mRNA with MEGAclearTM Kit (Ambion). .. The quality of the final product was confirmed by denaturing agarose electrophoresis.

    Purification:

    Article Title: Translational initiation factor eIF5 replaces eIF1 on the 40S ribosomal subunit to promote start-codon recognition
    Article Snippet: .. A single mRNA band, visualized by UV light, was excised from the gel and mRNA was electro-eluted in TBE buffer, concentrated and buffer exchanged by dialisis into storage buffer (10 mM ammonium acetate, pH 5.0, 50 mM KCl). mRNA was capped with Vaccinia Capping System (New England Biolabs, M2080S) according to the manufacturer’s instructions and further purified on an 8M Urea, 12% acrylamide gel as described above. .. The final concentration of mRNA was determined by A260 measurement.

    SYBR Green Assay:

    Article Title: La-related protein 1 (LARP1) repression of TOP mRNA translation is mediated through its cap-binding domain and controlled by an adjacent regulatory region
    Article Snippet: .. Materials Reagents were obtained from the following sources: antibodies for S6K, phospho-T389-S6K, eIF2α, phospho-Ser51-eIF2α, Raptor, mTOR, 4EBP1, LARP1, NCBP1, eIF4E, eIF4G and PABP from Cell Signaling Technology; primary antibodies for eIF3b and HRP-labeled secondary antibodies from Santa Cruz Biotechnology; IRDye secondary antibodies from LI-COR; Dulbecco’s modified Eagle’s medium (DMEM) from Life Technologies; heat-inactivated Fetal Bovine Serum (IFS) and 7mGDP from Sigma Aldrich; DNase I, T4 DNA ligase 1, T4 RNA Ligase I, T7 RNA polymerase, polynucleotide kinase, Protoscript II reverse transcriptase, Vaccinia Capping System, Oligo d(T)25 Magnetic beads and streptavidin-coated magnetic beads from New England Biolabs; iTaq Universal SYBR Green Supermix and Bradford Protein Assay from Bio-rad; RNeasy Plus Mini Kit from Qiagen; Dual Luciferase Assay from Promega; and X-tremeGENE 9 transfection reagent from Roche. .. Cell culture and preparation of cell extracts Cells were grown in high-glucose DMEM supplemented with 10% (v/v) heat-inactivated fetal bovine serum and penicillin-streptomycin.

    Modification:

    Article Title: La-related protein 1 (LARP1) repression of TOP mRNA translation is mediated through its cap-binding domain and controlled by an adjacent regulatory region
    Article Snippet: .. Materials Reagents were obtained from the following sources: antibodies for S6K, phospho-T389-S6K, eIF2α, phospho-Ser51-eIF2α, Raptor, mTOR, 4EBP1, LARP1, NCBP1, eIF4E, eIF4G and PABP from Cell Signaling Technology; primary antibodies for eIF3b and HRP-labeled secondary antibodies from Santa Cruz Biotechnology; IRDye secondary antibodies from LI-COR; Dulbecco’s modified Eagle’s medium (DMEM) from Life Technologies; heat-inactivated Fetal Bovine Serum (IFS) and 7mGDP from Sigma Aldrich; DNase I, T4 DNA ligase 1, T4 RNA Ligase I, T7 RNA polymerase, polynucleotide kinase, Protoscript II reverse transcriptase, Vaccinia Capping System, Oligo d(T)25 Magnetic beads and streptavidin-coated magnetic beads from New England Biolabs; iTaq Universal SYBR Green Supermix and Bradford Protein Assay from Bio-rad; RNeasy Plus Mini Kit from Qiagen; Dual Luciferase Assay from Promega; and X-tremeGENE 9 transfection reagent from Roche. .. Cell culture and preparation of cell extracts Cells were grown in high-glucose DMEM supplemented with 10% (v/v) heat-inactivated fetal bovine serum and penicillin-streptomycin.

    Incubation:

    Article Title: Elimination of Ribosome Inactivating Factors Improves the Efficiency of Bacillus subtilis and Saccharomyces cerevisiae Cell-Free Translation Systems
    Article Snippet: .. A typical 20 μL reaction containing 1 pmole DNA was incubated for 2 h at 37°C prior to mRNA isolation with MEGAclearTM Kit (Ambion), followed by capping by the Vaccinia Capping System (NEB) and re-purification of mRNA with MEGAclearTM Kit (Ambion). .. The quality of the final product was confirmed by denaturing agarose electrophoresis.

    Article Title: Correction of bleeding in experimental severe hemophilia A by systemic delivery of factor VIII-encoding mRNA
    Article Snippet: .. To obtain cap1, RNA was incubated with guanylyltransferase and 2 ′-O -methyltransferase (Vaccinia capping system; New England Biolabs, Frankfurt, Germany). .. All mRNA were purified and stored at −20°C.

    Bradford Protein Assay:

    Article Title: La-related protein 1 (LARP1) repression of TOP mRNA translation is mediated through its cap-binding domain and controlled by an adjacent regulatory region
    Article Snippet: .. Materials Reagents were obtained from the following sources: antibodies for S6K, phospho-T389-S6K, eIF2α, phospho-Ser51-eIF2α, Raptor, mTOR, 4EBP1, LARP1, NCBP1, eIF4E, eIF4G and PABP from Cell Signaling Technology; primary antibodies for eIF3b and HRP-labeled secondary antibodies from Santa Cruz Biotechnology; IRDye secondary antibodies from LI-COR; Dulbecco’s modified Eagle’s medium (DMEM) from Life Technologies; heat-inactivated Fetal Bovine Serum (IFS) and 7mGDP from Sigma Aldrich; DNase I, T4 DNA ligase 1, T4 RNA Ligase I, T7 RNA polymerase, polynucleotide kinase, Protoscript II reverse transcriptase, Vaccinia Capping System, Oligo d(T)25 Magnetic beads and streptavidin-coated magnetic beads from New England Biolabs; iTaq Universal SYBR Green Supermix and Bradford Protein Assay from Bio-rad; RNeasy Plus Mini Kit from Qiagen; Dual Luciferase Assay from Promega; and X-tremeGENE 9 transfection reagent from Roche. .. Cell culture and preparation of cell extracts Cells were grown in high-glucose DMEM supplemented with 10% (v/v) heat-inactivated fetal bovine serum and penicillin-streptomycin.

    Acrylamide Gel Assay:

    Article Title: Translational initiation factor eIF5 replaces eIF1 on the 40S ribosomal subunit to promote start-codon recognition
    Article Snippet: .. A single mRNA band, visualized by UV light, was excised from the gel and mRNA was electro-eluted in TBE buffer, concentrated and buffer exchanged by dialisis into storage buffer (10 mM ammonium acetate, pH 5.0, 50 mM KCl). mRNA was capped with Vaccinia Capping System (New England Biolabs, M2080S) according to the manufacturer’s instructions and further purified on an 8M Urea, 12% acrylamide gel as described above. .. The final concentration of mRNA was determined by A260 measurement.

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  • 99
    New England Biolabs vaccinia capping system
    Vaccinia Capping System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/vaccinia capping system/product/New England Biolabs
    Average 99 stars, based on 51 article reviews
    Price from $9.99 to $1999.99
    vaccinia capping system - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

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