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m6atp  (Jena Bioscience)


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    Structured Review

    Jena Bioscience m6atp
    The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.
    M6atp, supplied by Jena Bioscience, used in various techniques. Bioz Stars score: 93/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/m6atp/N6-Methyl-ATP/bio_rxiv__2025__11__25__690160-169-25-26
    Average 93 stars, based on 23 article reviews
    m6atp - by Bioz Stars, 2026-09
    93/100 stars

    Images

    1) Product Images from "Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation"

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation

    Journal: bioRxiv

    doi: 10.1101/2025.11.25.690160

    The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.
    Figure Legend Snippet: The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.

    Techniques Used: In Vitro, Pull Down Assay, Activity Assay, Two Tailed Test, Methylation, Isolation, Western Blot, Control, Luciferase

    The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.
    Figure Legend Snippet: The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

    Techniques Used: Luciferase, Activity Assay, In Vitro, Transfection, Ex Vivo, Two Tailed Test

    Related Articles

    In Vitro:

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation
    Article Snippet: .. In vitro transcription was performed using the T7-Scribe Standard RNA IVT Kit (CellScript, USA) according to the manufacturer’s instructions. m6A-Methylated mRNAs were generated by incorporating m6ATP (Jena Bioscience, Germany, NU-1101L) along with ATP at defined ratios. .. Capping and polyadenylation were conducted using the ScriptCapTM m7G Capping System and the A-PlusTM Poly(A) Polymerase Tailing Kit (CellScript, USA) following the manufacturer’s protocols.

    Article Title: m 6 A potentiates Sxl alternative pre-mRNA splicing for robust Drosophila sex determination.
    Article Snippet: Markers were generated by © 2016 Macmillan Publishers Limited, part of Springer Nature. .. All rights reserved. using in vitro transcripts with or without m6ATP (Jena Bioscience), which were then digested with RNase T1, kinased with PNK in the presence of [γ -32P]-ATP. ..

    Generated:

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation
    Article Snippet: .. In vitro transcription was performed using the T7-Scribe Standard RNA IVT Kit (CellScript, USA) according to the manufacturer’s instructions. m6A-Methylated mRNAs were generated by incorporating m6ATP (Jena Bioscience, Germany, NU-1101L) along with ATP at defined ratios. .. Capping and polyadenylation were conducted using the ScriptCapTM m7G Capping System and the A-PlusTM Poly(A) Polymerase Tailing Kit (CellScript, USA) following the manufacturer’s protocols.

    Modification:

    Article Title: Global RNA modifications to the MALAT1 triple helix differentially affect thermostability and weaken binding to METTL16.
    Article Snippet: .. Briefly, reactions used homemade T7 RNA polymerase and included 2 mM each of ATP, CTP, GTP and UTP (MilliporeSigma) for unmodified RNA and then substitution of the appropriate unmodified NTP for one of the modified nucleotides, m6ATP, TP and m1-TP (Jena Biosciences), for modified RNAs. .. After NAP10 column purification, RNA was quantified using a NanoDrop OneC instrument (Thermo Fisher Scientific) and the yields were 40-60% less than unmodified RNA.

    Raw Material:

    Article Title: Transferable deep learning enables identification of multiple types of RNA modifications using nanopore direct RNA sequencing
    Article Snippet: PCR products containing T7 promoter and rice full-length cDNA 524 sequences were used as templates for in vitro transcription by MEGAscript T7 Kit 525 (ThermoFisher, AM1333, USA) at 37°C for 2.5 hours to produce non-modified rice mRNAs 526 as control. .. At the same time, the ATP in the raw material was replaced with m1ATP (N1-527 Methyl-ATP – Solid, Jena Bioscience, NU-1027-1) or m6ATP (N6-Methyl-ATP, Jena 528 Bioscience, NU-1101S), or CTP was replaced with m5CTP (5-Methyl-CTP, APExBio, B7967) 529 for producing m1A-modiifed RNAs, m6A-modfiied RNA and m5C-modified mRNAs, 530 respectively. ..

    Article Title: Transfer learning enables identification of multiple types of RNA modifications using nanopore direct RNA sequencing.
    Article Snippet: PCR products containing T7 promoter and rice cDNA sequences were used as templates for in vitro transcription by MEGAscript T7 Kit (ThermoFisher, AM1333, USA) at 37 °C for 2.5 h to produce non-modified rice mRNAs as control. .. At the same time, the ATP in the raw material was replaced with m1ATP (N1-Methyl-ATP – Solid, Jena Bioscience, NU-1027-1) or m6ATP (N6-Methyl-ATP, Jena Bioscience, NU-1101S), or CTPwas replacedwithm5CTP (5-Methyl-CTP, APExBio, B7967) for producing m1A-modiifed RNAs, m6A-modfiied RNA and m5C-modified mRNAs, respectively. ..



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    Image Search Results


    The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.

    Journal: bioRxiv

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation

    doi: 10.1101/2025.11.25.690160

    Figure Lengend Snippet: The NlucP reporter mRNAs with 5’ and 3’ UTRs of SLU7 , RPL32, and HSP70 were transcribed in vitro in the presence of 50% or 0% m⁶ATP. mRNAs were then enzymatically capped and polyadenylated (A-B) . For the pull-down assay (C) , after transcription, the mRNAs were first polyadenylated, then biotinylated at the 3’ end, and enzymatically capped. (A) mRNAs were translated in CFTS prepared from untreated HEK293T cells in the presence of 0.1 mM cap-analogs m7GpppG or ApppG. NlucP activity in the presence of m7GpppG was normalized to that in the presence of ApppG. (B) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in Torin1-treated CFTS was normalized to that in DMSO-treated CFTS. Values are the means of at least three independent replicates. Error bars show standard deviations. Two-tailed Student’s t-test was used to estimate the statistical significance between methylated and unmethylated mRNAs. ***p<0.001. (C) The biotinylated mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. RNA-protein complexes were isolated using streptavidin-sepharose and analysed by Western blot using antibodies against eIF4A, eIF4E, and eIF4G. Uncapped mRNA with RPL32 UTRs (K-) was used as a control. CFTS: cell-free translation system; RLU: relative luciferase units.

    Article Snippet: In vitro transcription was performed using the T7-Scribe Standard RNA IVT Kit (CellScript, USA) according to the manufacturer’s instructions. m6A-Methylated mRNAs were generated by incorporating m6ATP (Jena Bioscience, Germany, NU-1101L) along with ATP at defined ratios.

    Techniques: In Vitro, Pull Down Assay, Activity Assay, Two Tailed Test, Methylation, Isolation, Western Blot, Control, Luciferase

    The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

    Journal: bioRxiv

    Article Title: Transcript-wide m 6 A methylation defines the efficiency of the cap-independent translation initiation

    doi: 10.1101/2025.11.25.690160

    Figure Lengend Snippet: The barplots show the relative luciferase activity for the RPL32 reporter mRNAs transcribed in vitro with the indicated m⁶ATP percentage (X-axis). (A) mRNAs were translated in CFTS prepared from HEK293T cells treated with 250nM Torin1 or DMSO. NlucP activity in CFTS obtained from Torin1-treated cells was normalized to that in the CFTS from DMSO-treated cells. (B) mRNA transfected into WT HEK293T cells, pre-treated for 2h by 250nM Torin1 or DMSO. The mRNAs were transfected together with furimazine, and luciferase activity was measured in living cells at 40 min for the CFTS assay and at 80 min for the ex vivo assay. NlucP activity in Torin1-treated cells was normalized to that in DMSO-treated cells. Values are the means of at least three independent replicates. The error bars show standard deviations. The results were analyzed using the two-tailed Student’s t-test, with the significance levels indicated as follows: *p < 0.05; **p < 0.01; ***p < 0.001.

    Article Snippet: In vitro transcription was performed using the T7-Scribe Standard RNA IVT Kit (CellScript, USA) according to the manufacturer’s instructions. m6A-Methylated mRNAs were generated by incorporating m6ATP (Jena Bioscience, Germany, NU-1101L) along with ATP at defined ratios.

    Techniques: Luciferase, Activity Assay, In Vitro, Transfection, Ex Vivo, Two Tailed Test

    Journal: iScience

    Article Title: Distinct pathways utilized by METTL3 to regulate antiviral innate immune response

    doi: 10.1016/j.isci.2024.111071

    Figure Lengend Snippet:

    Article Snippet: EV71 genomic RNAs were in vitro transcribed from a HindIII (ThermoFisher)-linearized infectious DNA construct with ATP (ThermoFisher) or m6ATP (TriLink, San Diego, CA) as substrates using a MEGAscript T7 Kit (Ambion, Austin, TX, USA) and the RNAs were transfected into HEK293T cells using DMRIE-C Reagent (ThermoFisher).

    Techniques: Virus, Recombinant, Transfection, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Luciferase, SYBR Green Assay, Isolation, shRNA, Plasmid Preparation, Software