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monarch spin rna cleanup kit  (New England Biolabs)


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    New England Biolabs monarch spin rna cleanup kit
    Monarch Spin Rna Cleanup Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 423 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+cleanup+kit/Monarch+Spin+RNA+Cleanup+Kit/pmc13010419-15-0-6
    Average 99 stars, based on 423 article reviews
    monarch spin rna cleanup kit - by Bioz Stars, 2026-09
    99/100 stars

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    Related Articles

    Incubation:

    Article Title: Thioester-Containing Ionizable Lipids with Enhanced Endosomal Escape and Biodegradability for mRNA and tRNA Delivery
    Article Snippet: The resulting mRNA was purified using an Oligo-dT resin column on an ÄKTA chromatography system (Cytiva, Marlborough, MA, USA), following the manufacturer’s protocol, yielding highly pure transcripts suitable for downstream applications. tRNA was synthesized in vitro using a specifically designed dsDNA (GenScript, Piscataway, NJ, USA) template rather than a linearized plasmid. .. The IVT reaction was similar to that used for mRNA, with the following modifications: UTP (5 μg/mL; Hongene, R5331) was used instead of N1-methylpseudouridine, CleanCap® AG was omitted, and the reaction was incubated for 5 h to optimize tRNA yield. tRNA was subsequently purified using the Monarch® Spin RNA Cleanup Kit (50 μg; NEB, Ipswich, MA, USA) according to the manufacturer’s protocol, effectively removing residual enzymes and unincorporated nucleotides. ..

    Purification:

    Article Title: Thioester-Containing Ionizable Lipids with Enhanced Endosomal Escape and Biodegradability for mRNA and tRNA Delivery
    Article Snippet: The resulting mRNA was purified using an Oligo-dT resin column on an ÄKTA chromatography system (Cytiva, Marlborough, MA, USA), following the manufacturer’s protocol, yielding highly pure transcripts suitable for downstream applications. tRNA was synthesized in vitro using a specifically designed dsDNA (GenScript, Piscataway, NJ, USA) template rather than a linearized plasmid. .. The IVT reaction was similar to that used for mRNA, with the following modifications: UTP (5 μg/mL; Hongene, R5331) was used instead of N1-methylpseudouridine, CleanCap® AG was omitted, and the reaction was incubated for 5 h to optimize tRNA yield. tRNA was subsequently purified using the Monarch® Spin RNA Cleanup Kit (50 μg; NEB, Ipswich, MA, USA) according to the manufacturer’s protocol, effectively removing residual enzymes and unincorporated nucleotides. ..

    Article Title: ACRC/GCNA is an essential protease that repairs DNA–protein crosslinks during vertebrate development
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence ( NM_052957.5 ) from a plasmid we obtained from the Niels Mailand’s group (HsACRC-pAcGFP1-C1) [ ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1–2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/μl RNA). ..

    Article Title: Human PARPs modify RNA nucleobases in vitro and in cells
    Article Snippet: Following the reaction, proteins were digested with 20 U Proteinase K (New England Biolabs) for 20 min at 37°C, and samples were resolved on denaturing urea-PAGE. .. For ADP-ribosylhydrolase assays, RNA oligonucleotides were first modified in 100-200 μL ADP-ribosylation reactions, then purified using the Monarch® Spin RNA Cleanup Kit (New England Biolabs) before use in hydrolase reactions. ..

    Article Title: ACRC/GCNA is an essential protease that repairs DNA-protein crosslinks during vertebrate development.
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence (NM_052957.5) from a plasmid we obtained from the Niels Mailand’s group (HsACRCpAcGFP1-C1) [ 23 ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1 –2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/ μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/ μl RNA). ..

    Article Title: Integrated Size-Selective Cell Purification and Electroporation for Genetic Manipulation of Primary Cells
    Article Snippet: A Poly-(A) tail of approximately 100 nt was added using E. coli Poly(A) Polymerase (cat#M0276L, New England Biolabs, This page contains the following errors:) to the mRNA product. .. After each step, RNA products were purified using an RNA Cleanup Kit (cat# T2050S, New England Biolabs, Ipswich, MA, USA). .. RNA concentration and purity were assessed using a Nanodrop spectrophotometer.

    Article Title: Transcriptomic and RNA Modification Landscape of Severe Fever with Thrombocytopenia Syndrome Virus Revealed by Nanopore Direct RNA Sequencing.
    Article Snippet: The PCR products were purified using an agarose gel electrophoresis purification kit (Wizard® SV Gel and PCR Clean-Up System, Promega, Madison, WI, USA, A9282), followed by in vitro transcription using the T7 High Yield RNA Transcription Kit (Vazyme, Nanjing, China, TR101). .. Finally, the resulting RNA products were purified using the Monarch® Spin RNA Cleanup Kit (NEB, Ipswich, MA, USA, T2050L). ..

    Article Title: Integrated Size-Selective Cell Purification and Electroporation for Genetic Manipulation of Primary Cells
    Article Snippet: A Poly-(A) tail of approximately 100 nt was added using E. coli Poly(A) Polymerase (cat#M0276L, New England Biolabs, This page contains the following errors:) to the mRNA product. .. After each step, RNA products were purified using an RNA Cleanup Kit (cat# T2050S, New England Biolabs, Ipswich, MA, USA). .. RNA concentration and purity were assessed using a Nanodrop spectrophotometer.

    Injection:

    Article Title: ACRC/GCNA is an essential protease that repairs DNA–protein crosslinks during vertebrate development
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence ( NM_052957.5 ) from a plasmid we obtained from the Niels Mailand’s group (HsACRC-pAcGFP1-C1) [ ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1–2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/μl RNA). ..

    Article Title: ACRC/GCNA is an essential protease that repairs DNA-protein crosslinks during vertebrate development.
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence (NM_052957.5) from a plasmid we obtained from the Niels Mailand’s group (HsACRCpAcGFP1-C1) [ 23 ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1 –2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/ μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/ μl RNA). ..

    Mutagenesis:

    Article Title: ACRC/GCNA is an essential protease that repairs DNA–protein crosslinks during vertebrate development
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence ( NM_052957.5 ) from a plasmid we obtained from the Niels Mailand’s group (HsACRC-pAcGFP1-C1) [ ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1–2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/μl RNA). ..

    Article Title: ACRC/GCNA is an essential protease that repairs DNA-protein crosslinks during vertebrate development.
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence (NM_052957.5) from a plasmid we obtained from the Niels Mailand’s group (HsACRCpAcGFP1-C1) [ 23 ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1 –2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/ μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/ μl RNA). ..

    Control:

    Article Title: ACRC/GCNA is an essential protease that repairs DNA–protein crosslinks during vertebrate development
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence ( NM_052957.5 ) from a plasmid we obtained from the Niels Mailand’s group (HsACRC-pAcGFP1-C1) [ ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1–2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/μl RNA). ..

    Article Title: ACRC/GCNA is an essential protease that repairs DNA-protein crosslinks during vertebrate development.
    Article Snippet: We have sub-cloned the full-length human ACRC coding sequence (NM_052957.5) from a plasmid we obtained from the Niels Mailand’s group (HsACRCpAcGFP1-C1) [ 23 ] into pCS2 + HisMyc vector. mRNA was synthesized using purified, linearized plasmid by performing an in vitro transcription with SP6 using the Hiscribe SP6 RNA kit (#E2070S, NEB) and a cap analog from the ARCA kit (#S1411, NEB) for improved mRNA stability. .. The resulting mRNA was purified using the RNA cleanup kit (#T2040, NEB) and injected into 1 –2 cell stage acrc maternal mutant embryos or into control WT embryos (1 nl of the mRNA solution: 250 ng/ μl RNA in 0.3M KCl and 0.015% phenol red, except for DrSprtn: 100 ng/ μl RNA). ..

    Modification:

    Article Title: Human PARPs modify RNA nucleobases in vitro and in cells
    Article Snippet: Following the reaction, proteins were digested with 20 U Proteinase K (New England Biolabs) for 20 min at 37°C, and samples were resolved on denaturing urea-PAGE. .. For ADP-ribosylhydrolase assays, RNA oligonucleotides were first modified in 100-200 μL ADP-ribosylation reactions, then purified using the Monarch® Spin RNA Cleanup Kit (New England Biolabs) before use in hydrolase reactions. ..



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    Purification of circRNAs and extending the <t>RNA</t> ligase 2 (RL2)-dependent circularization method to other RNAs. ( A ) CircRNAs synthesized with RNA ligase 2 using DNA splint <t>were</t> <t>purified</t> using three different approaches: from 3% urea–PAGE using crush and soak method, or from EX E-gels either using the crush and soak method or using column-based kit. As a control, linear RNAs were also extracted using the same methods. CircRNAs extracted from 3% urea–PAGE or EX E-gel using a crush and soak method had more intact circRNAs with less nicking compared to those extracted from EX E-gel using column-based kits. Linear RNAs on the other hand remained intact with each of the approaches. ( B ) Schematic of RNase-H based circularity confirmation assay that uses a short ssDNA probe which cleaves intact circRNAs into a single linear band, while nicked circRNAs or linear RNAs are cut into two shorter bands. ( C ) RNase-H based assay confirmed circularity of EGFP-IRES circRNAs. Linear RNAs were cleaved into two shorter bands of expected sizes while circRNAs derived from modified DNA templates were linearized to the size of full-length linear precursor. ( D, E ) 5′-monophosphate linear precursors of human immunodeficiency virus (HIV) and mCherry were ligated using RNA ligase 2 and respective DNA splints. For circHIV, urea–PAGE purification of circRNAs derived from modified templates had the highest yields with the least contaminating RNAs. Yields of mCherry circRNAs were much higher with polyA + RNase R approach on RNAs from modified template ligated using RNA ligase 2, however urea–PAGE showed higher and lower sized undesired RNAs. Representative data are from a mean of n = 3 technical replicates with SEM. ( F ) Sanger sequencing confirmed accuracy of circRNAs. Clean chromatograms were observed for ligation junctions of both HlV and mCherry circRNAs derived from modified DNA templates purified either through poly(A) tailing and RNase R treatment or from urea–PAGE purification.
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    Image Search Results


    Purification of circRNAs and extending the RNA ligase 2 (RL2)-dependent circularization method to other RNAs. ( A ) CircRNAs synthesized with RNA ligase 2 using DNA splint were purified using three different approaches: from 3% urea–PAGE using crush and soak method, or from EX E-gels either using the crush and soak method or using column-based kit. As a control, linear RNAs were also extracted using the same methods. CircRNAs extracted from 3% urea–PAGE or EX E-gel using a crush and soak method had more intact circRNAs with less nicking compared to those extracted from EX E-gel using column-based kits. Linear RNAs on the other hand remained intact with each of the approaches. ( B ) Schematic of RNase-H based circularity confirmation assay that uses a short ssDNA probe which cleaves intact circRNAs into a single linear band, while nicked circRNAs or linear RNAs are cut into two shorter bands. ( C ) RNase-H based assay confirmed circularity of EGFP-IRES circRNAs. Linear RNAs were cleaved into two shorter bands of expected sizes while circRNAs derived from modified DNA templates were linearized to the size of full-length linear precursor. ( D, E ) 5′-monophosphate linear precursors of human immunodeficiency virus (HIV) and mCherry were ligated using RNA ligase 2 and respective DNA splints. For circHIV, urea–PAGE purification of circRNAs derived from modified templates had the highest yields with the least contaminating RNAs. Yields of mCherry circRNAs were much higher with polyA + RNase R approach on RNAs from modified template ligated using RNA ligase 2, however urea–PAGE showed higher and lower sized undesired RNAs. Representative data are from a mean of n = 3 technical replicates with SEM. ( F ) Sanger sequencing confirmed accuracy of circRNAs. Clean chromatograms were observed for ligation junctions of both HlV and mCherry circRNAs derived from modified DNA templates purified either through poly(A) tailing and RNase R treatment or from urea–PAGE purification.

    Journal: Nucleic Acids Research

    Article Title: Generation of precise and accurate engineered circRNAs using enzymatic ligation

    doi: 10.1093/nar/gkag405

    Figure Lengend Snippet: Purification of circRNAs and extending the RNA ligase 2 (RL2)-dependent circularization method to other RNAs. ( A ) CircRNAs synthesized with RNA ligase 2 using DNA splint were purified using three different approaches: from 3% urea–PAGE using crush and soak method, or from EX E-gels either using the crush and soak method or using column-based kit. As a control, linear RNAs were also extracted using the same methods. CircRNAs extracted from 3% urea–PAGE or EX E-gel using a crush and soak method had more intact circRNAs with less nicking compared to those extracted from EX E-gel using column-based kits. Linear RNAs on the other hand remained intact with each of the approaches. ( B ) Schematic of RNase-H based circularity confirmation assay that uses a short ssDNA probe which cleaves intact circRNAs into a single linear band, while nicked circRNAs or linear RNAs are cut into two shorter bands. ( C ) RNase-H based assay confirmed circularity of EGFP-IRES circRNAs. Linear RNAs were cleaved into two shorter bands of expected sizes while circRNAs derived from modified DNA templates were linearized to the size of full-length linear precursor. ( D, E ) 5′-monophosphate linear precursors of human immunodeficiency virus (HIV) and mCherry were ligated using RNA ligase 2 and respective DNA splints. For circHIV, urea–PAGE purification of circRNAs derived from modified templates had the highest yields with the least contaminating RNAs. Yields of mCherry circRNAs were much higher with polyA + RNase R approach on RNAs from modified template ligated using RNA ligase 2, however urea–PAGE showed higher and lower sized undesired RNAs. Representative data are from a mean of n = 3 technical replicates with SEM. ( F ) Sanger sequencing confirmed accuracy of circRNAs. Clean chromatograms were observed for ligation junctions of both HlV and mCherry circRNAs derived from modified DNA templates purified either through poly(A) tailing and RNase R treatment or from urea–PAGE purification.

    Article Snippet: The final reactions from all the approaches were purified using RNA cleanup columns (NEB #T2030L).

    Techniques: Purification, Synthesized, Control, Rnase H Assay, Derivative Assay, Modification, Virus, Sequencing, Ligation