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monarch rna cleanup binding buffer  (New England Biolabs)


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    New England Biolabs monarch rna cleanup binding buffer
    In vitro hypoxia/reperfusion (iH/R) stress regulates m6A levels of key anti-apoptotic and pro-apoptotic genes. ( A–C ). Ctrl and Mettl3 mofified HL1 cardiomyocytes were subjected to sham or iH/R, and total <t>RNA</t> was subjected to <t>m6A</t> <t>RIP,</t> followed by RT-qPCR using primers for the indicated pro-apoptotic genes (Bax and PTEN) and anti-apoptitic gene (Bcl2). Values are normalized to input. n = 3–5, * p < 0.05; ** p < 0.01; *** p < 0.001.
    Monarch Rna Cleanup Binding Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 92/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/rna+binding+buffer/Monarch+RNA+Cleanup+Binding+Buffer/pmc08369344-86-60-70
    Average 92 stars, based on 31 article reviews
    monarch rna cleanup binding buffer - by Bioz Stars, 2026-09
    92/100 stars

    Images

    1) Product Images from "Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice"

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice

    Journal: Frontiers in Pharmacology

    doi: 10.3389/fphar.2021.654316

    In vitro hypoxia/reperfusion (iH/R) stress regulates m6A levels of key anti-apoptotic and pro-apoptotic genes. ( A–C ). Ctrl and Mettl3 mofified HL1 cardiomyocytes were subjected to sham or iH/R, and total RNA was subjected to m6A RIP, followed by RT-qPCR using primers for the indicated pro-apoptotic genes (Bax and PTEN) and anti-apoptitic gene (Bcl2). Values are normalized to input. n = 3–5, * p < 0.05; ** p < 0.01; *** p < 0.001.
    Figure Legend Snippet: In vitro hypoxia/reperfusion (iH/R) stress regulates m6A levels of key anti-apoptotic and pro-apoptotic genes. ( A–C ). Ctrl and Mettl3 mofified HL1 cardiomyocytes were subjected to sham or iH/R, and total RNA was subjected to m6A RIP, followed by RT-qPCR using primers for the indicated pro-apoptotic genes (Bax and PTEN) and anti-apoptitic gene (Bcl2). Values are normalized to input. n = 3–5, * p < 0.05; ** p < 0.01; *** p < 0.001.

    Techniques Used: In Vitro, Quantitative RT-PCR

    Related Articles

    Plasmid Preparation:

    Article Title: The retinoic acid family-like nuclear receptor SmRAR identified by single-cell transcriptomics of ovarian cells controls oocyte differentiation in Schistosoma mansoni.
    Article Snippet: .. For this, 1 μg of pJC53.2 plasmid DNA was digested by Ahd I (New England Biolabs; NEB, UK) in a total volume of 50 μl containing 1 × CutSmart buffer (NEB, UK) at 37 ◦C for 2 h. The resulting DNA fragments were separated by agarose gel electrophoresis and extracted using the Monarch DNA Cleanup and Gel Extraction Kit (NEB, UK). .. Amplicons of 500 bp were obtained by polymerase chain reaction (PCR), using a final reaction volume of 20 μl, including 100 ng S. mansoni cDNA generated by reverse transcription of extracted total RNA of couples (QuantiTect Reverse Transcription Kit, Qiagen, Hilden, Germany).

    Article Title: The retinoic acid family-like nuclear receptor SmRAR identified by single-cell transcriptomics of ovarian cells controls oocyte differentiation in Schistosoma mansoni
    Article Snippet: .. For this, 1 μg of pJC53.2 plasmid DNA was digested by Ahd I (New England Biolabs; NEB, UK) in a total volume of 50 μl containing 1× CutSmart buffer (NEB, UK) at 37°C for 2 h. The resulting DNA fragments were separated by agarose gel electrophoresis and extracted using the Monarch DNA Cleanup and Gel Extraction Kit (NEB, UK). .. Amplicons of 500 bp were obtained by polymerase chain reaction (PCR), using a final reaction volume of 20 μl, including 100 ng S. mansoni cDNA generated by reverse transcription of extracted total RNA of couples (QuantiTect Reverse Transcription Kit, Qiagen, Hilden, Germany).

    Agarose Gel Electrophoresis:

    Article Title: The retinoic acid family-like nuclear receptor SmRAR identified by single-cell transcriptomics of ovarian cells controls oocyte differentiation in Schistosoma mansoni.
    Article Snippet: .. For this, 1 μg of pJC53.2 plasmid DNA was digested by Ahd I (New England Biolabs; NEB, UK) in a total volume of 50 μl containing 1 × CutSmart buffer (NEB, UK) at 37 ◦C for 2 h. The resulting DNA fragments were separated by agarose gel electrophoresis and extracted using the Monarch DNA Cleanup and Gel Extraction Kit (NEB, UK). .. Amplicons of 500 bp were obtained by polymerase chain reaction (PCR), using a final reaction volume of 20 μl, including 100 ng S. mansoni cDNA generated by reverse transcription of extracted total RNA of couples (QuantiTect Reverse Transcription Kit, Qiagen, Hilden, Germany).

    Article Title: The retinoic acid family-like nuclear receptor SmRAR identified by single-cell transcriptomics of ovarian cells controls oocyte differentiation in Schistosoma mansoni
    Article Snippet: .. For this, 1 μg of pJC53.2 plasmid DNA was digested by Ahd I (New England Biolabs; NEB, UK) in a total volume of 50 μl containing 1× CutSmart buffer (NEB, UK) at 37°C for 2 h. The resulting DNA fragments were separated by agarose gel electrophoresis and extracted using the Monarch DNA Cleanup and Gel Extraction Kit (NEB, UK). .. Amplicons of 500 bp were obtained by polymerase chain reaction (PCR), using a final reaction volume of 20 μl, including 100 ng S. mansoni cDNA generated by reverse transcription of extracted total RNA of couples (QuantiTect Reverse Transcription Kit, Qiagen, Hilden, Germany).

    Gel Extraction:

    Article Title: The retinoic acid family-like nuclear receptor SmRAR identified by single-cell transcriptomics of ovarian cells controls oocyte differentiation in Schistosoma mansoni.
    Article Snippet: .. For this, 1 μg of pJC53.2 plasmid DNA was digested by Ahd I (New England Biolabs; NEB, UK) in a total volume of 50 μl containing 1 × CutSmart buffer (NEB, UK) at 37 ◦C for 2 h. The resulting DNA fragments were separated by agarose gel electrophoresis and extracted using the Monarch DNA Cleanup and Gel Extraction Kit (NEB, UK). .. Amplicons of 500 bp were obtained by polymerase chain reaction (PCR), using a final reaction volume of 20 μl, including 100 ng S. mansoni cDNA generated by reverse transcription of extracted total RNA of couples (QuantiTect Reverse Transcription Kit, Qiagen, Hilden, Germany).

    Article Title: The retinoic acid family-like nuclear receptor SmRAR identified by single-cell transcriptomics of ovarian cells controls oocyte differentiation in Schistosoma mansoni
    Article Snippet: .. For this, 1 μg of pJC53.2 plasmid DNA was digested by Ahd I (New England Biolabs; NEB, UK) in a total volume of 50 μl containing 1× CutSmart buffer (NEB, UK) at 37°C for 2 h. The resulting DNA fragments were separated by agarose gel electrophoresis and extracted using the Monarch DNA Cleanup and Gel Extraction Kit (NEB, UK). .. Amplicons of 500 bp were obtained by polymerase chain reaction (PCR), using a final reaction volume of 20 μl, including 100 ng S. mansoni cDNA generated by reverse transcription of extracted total RNA of couples (QuantiTect Reverse Transcription Kit, Qiagen, Hilden, Germany).

    Polymerase Chain Reaction:

    Article Title: Whole-genome functional characterization of RE1 silencers using a modified massively parallel reporter assay.
    Article Snippet: A GFP open reading frame (ORF) with a minimal promoter and partial 30 UTR was amplified from pMPRAduo:minP:GFP (addgene pending) in 1600 mL volume containing 800 mL of Q5 High-Fidelity 2X Master Mix (NEB, M0492L) and 0.5 mM forward and reverse primers (primers 7 and 8 for vector A and primers 9 and 10 for vector P, Table S1) cycled with the following conditions: 98 C for 30s, 20 cycles of (98 C for 10 s, 60 C for 15 s, 72 C 45 s), 72 C for 5 min. .. The PCR product was purified using 1.53 volume of AMpure XP and inserted by gibson assembly using 3 mg of linearized Dorf library and 9 mg of the GFP amplicon in 100 mL total volume for 90min at 50 C. Assembled vectors were purified using 13 volume of AMpure XP and a secondary digest performed with 40 U of AsiSI or PmeI, 5 U of RecBCD (NEB, M0345), 10 mg BSA, 1 mM ATP, 13 NEB Buffer 4 at 37 C overnight followed by purification with a Monarch PCR & DNA Clean up kit using 12 mL of water for elution. ..

    Purification:

    Article Title: Whole-genome functional characterization of RE1 silencers using a modified massively parallel reporter assay.
    Article Snippet: A GFP open reading frame (ORF) with a minimal promoter and partial 30 UTR was amplified from pMPRAduo:minP:GFP (addgene pending) in 1600 mL volume containing 800 mL of Q5 High-Fidelity 2X Master Mix (NEB, M0492L) and 0.5 mM forward and reverse primers (primers 7 and 8 for vector A and primers 9 and 10 for vector P, Table S1) cycled with the following conditions: 98 C for 30s, 20 cycles of (98 C for 10 s, 60 C for 15 s, 72 C 45 s), 72 C for 5 min. .. The PCR product was purified using 1.53 volume of AMpure XP and inserted by gibson assembly using 3 mg of linearized Dorf library and 9 mg of the GFP amplicon in 100 mL total volume for 90min at 50 C. Assembled vectors were purified using 13 volume of AMpure XP and a secondary digest performed with 40 U of AsiSI or PmeI, 5 U of RecBCD (NEB, M0345), 10 mg BSA, 1 mM ATP, 13 NEB Buffer 4 at 37 C overnight followed by purification with a Monarch PCR & DNA Clean up kit using 12 mL of water for elution. ..

    Amplification:

    Article Title: Whole-genome functional characterization of RE1 silencers using a modified massively parallel reporter assay.
    Article Snippet: A GFP open reading frame (ORF) with a minimal promoter and partial 30 UTR was amplified from pMPRAduo:minP:GFP (addgene pending) in 1600 mL volume containing 800 mL of Q5 High-Fidelity 2X Master Mix (NEB, M0492L) and 0.5 mM forward and reverse primers (primers 7 and 8 for vector A and primers 9 and 10 for vector P, Table S1) cycled with the following conditions: 98 C for 30s, 20 cycles of (98 C for 10 s, 60 C for 15 s, 72 C 45 s), 72 C for 5 min. .. The PCR product was purified using 1.53 volume of AMpure XP and inserted by gibson assembly using 3 mg of linearized Dorf library and 9 mg of the GFP amplicon in 100 mL total volume for 90min at 50 C. Assembled vectors were purified using 13 volume of AMpure XP and a secondary digest performed with 40 U of AsiSI or PmeI, 5 U of RecBCD (NEB, M0345), 10 mg BSA, 1 mM ATP, 13 NEB Buffer 4 at 37 C overnight followed by purification with a Monarch PCR & DNA Clean up kit using 12 mL of water for elution. ..

    Magnetic Beads:

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice
    Article Snippet: .. Briefly, 2 μl N6-Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIP-enriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice.
    Article Snippet: .. Briefly, 2 μl N6Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIPenriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..

    Incubation:

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice
    Article Snippet: .. Briefly, 2 μl N6-Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIP-enriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice.
    Article Snippet: .. Briefly, 2 μl N6Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIPenriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..

    Isolation:

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice
    Article Snippet: .. Briefly, 2 μl N6-Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIP-enriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice.
    Article Snippet: .. Briefly, 2 μl N6Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIPenriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..

    Binding Assay:

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice
    Article Snippet: .. Briefly, 2 μl N6-Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIP-enriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..

    Article Title: Aging-Associated Differences in Epitranscriptomic m6A Regulation in Response to Acute Cardiac Ischemia/Reperfusion Injury in Female Mice.
    Article Snippet: .. Briefly, 2 μl N6Methyladenosine Antibody was added to protein G magnetic beads (Thermo Fisher) and incubated at 4°C for 2 h. Following two washes in reaction buffer, the RNA was added to the antibody–bead mixture containing RNasin Plus RNase Inhibitor (Promega) and incubated at 4°C for 2 h. Next, RIPenriched RNAs were isolated from the antibody-immobilized protein G beads using Monarch RNA cleanup binding buffer from Monarch RNA Cleanup kit (New England Biolabs). ..



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