3 o me m7g  (New England Biolabs)


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    Name:
    3 O Me m7G 5 ppp 5 G RNA Cap Structure Analog
    Description:
    3 O Me m7G 5 ppp 5 G RNA Cap Structure Analog 5 umol
    Catalog Number:
    s1411l
    Price:
    604
    Size:
    5 umol
    Category:
    Capping Reagents for DNA RNA Synthesis
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    Structured Review

    New England Biolabs 3 o me m7g
    3 O Me m7G 5 ppp 5 G RNA Cap Structure Analog
    3 O Me m7G 5 ppp 5 G RNA Cap Structure Analog 5 umol
    https://www.bioz.com/result/3 o me m7g/product/New England Biolabs
    Average 95 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    3 o me m7g - by Bioz Stars, 2020-02
    95/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Generation and Characterization of Closely Related Epizootic and Enzootic Infectious cDNA Clones for Studying Interferon Sensitivity and Emergence Mechanisms of Venezuelan Equine Encephalitis Virus
    Article Snippet: Both cDNA clones were purified by using the Qiagen MiniPrep kit. .. RNAs were transcribed in the presence of m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs) by using SP6 RNA polymerase (Invitrogen).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: Viral translation assay A DENV-2 reporter construct was generated by cloning T7 promoter, DENV-2 5′-UTR, the first 72 nucleotides of DENV-2 capsid-coding sequence, firefly luciferase gene and DENV-2 3′-UTR into pGL3-Basic (Promega) according to a previously described method [ ]. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    Amplification:

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: PCR products were generated using the following amplification protocol: initial denaturation step at 94°C for 3 min, followed by 25 cycles of denaturation at 94°C for 45 s, annealing at 57°C for 1 min, extension at 72°C for 1 min, and final extension at 72°C for 5 min. .. Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: Briefly, pcDNA 3.3 plasmid (Aldevron, Fargo, ND, USA) insert encoding eGFP and pEX-K4 plasmid insert encoding Link N (Eurofins Genomics GmbH, Ebersberg, Germany) were amplified using the Hot Star HiFidelity Polymerase Kit (Qiagen, Hilden, Germany), according to manufacturer’s instructions. .. IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany).

    Reporter Assay:

    Article Title: Adaptor Protein 1A Facilitates Dengue Virus Replication
    Article Snippet: One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. Following 8 h after transfection with reporter RNA, cells were harvested and determined for Renilla luciferase expression using Luciferase Reporter Assay System (Promega).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. Following 12 h after transfection with viral and control reporter RNA, cells were harvested and determined for firefly and Renilla luciferase expression using Dual-Luciferase Reporter Assay System (Promega).

    Expressing:

    Article Title: Adaptor Protein 1A Facilitates Dengue Virus Replication
    Article Snippet: One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. Following 8 h after transfection with reporter RNA, cells were harvested and determined for Renilla luciferase expression using Luciferase Reporter Assay System (Promega).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. Following 12 h after transfection with viral and control reporter RNA, cells were harvested and determined for firefly and Renilla luciferase expression using Dual-Luciferase Reporter Assay System (Promega).

    Article Title: Optimized conditions for successful transfection of human endothelial cells with in vitro synthesized and modified mRNA for induction of protein expression
    Article Snippet: The generated mRNA transcript is then used to induce protein expression in cells. .. At first, 23 μl NTP/cap analog mixture containing 7.5 mM ATP, 1.875 mM GTP (both from MEGAscript® T7 Kit), 7.5 mM Me-CTP, 7.5 mM Pseudo-UTP (both from TriLink BioTechnologies, San Diego, USA), and 2.5 mM 3′-O-Me-m7 G(5′)ppp(5′)G RNA cap structure analog (New England Biolabs, Frankfurt am Main, Germany) was prepared and mixed thoroughly.

    Synthesized:

    Article Title: Dom34 mediates targeting of exogenous RNA in the antiviral OAS/RNase L pathway
    Article Snippet: 5×Flag-EGFP mRNA, EGFP (1–87) mRNA control and EGFP (1–87) mRNA AUG mt were synthesized using T7 RNA polymerase (TAKARA) according to the manufacturer's instructions. pBK-5F-EGFP, pBK-EGFP (1–87) control and pBK-EGFP (1–87) ATG mt that were linearized by BsmBI were used as templates for the in vitro synthesis of 5×Flag-EGFP, EGFP (1–87) mRNA control and EGFP (1–87) mRNA AUG mt, respectively. .. For the synthesis of mRNA whose 5′-terminus was modified with a cap structure, 3′-O-Me-m7G(5′)ppp(5′)G cap analog (New England Biolabs) was added during the T7 polymerase reaction.

    Article Title: Combination Immunotherapy of MUC1 mRNA Nano-vaccine and CTLA-4 Blockade Effectively Inhibits Growth of Triple Negative Breast Cancer
    Article Snippet: Mannose-PEG-DSPE was synthesized using DSPE-PEG-NHS and 4-amino phenyl-mannopyranoside (Sigma-Aldrich, St. Louis, MO) according to the previously established protocol in our laboratory. .. 3′-O-Me-m7 G (5′) ppp (5′) G RNA cap structure analog was purchased from NEB.

    Article Title: Generation and Functional In Vitro Analysis of Semliki Forest Virus Vectors Encoding TNF-α and IFN-γ
    Article Snippet: RNA Synthesis and Production of Viral Particles RNAs were synthesized and transfected into BHK-21 cells using electroporation as previously described ( ). .. The transcribed rRNAs were capped by adding 1 mM 3′-O -Me-m7 G(5′)ppp(5′)G cap-structure analog (Cat. No. S1411S; New England Biolabs, Hitchin, UK) during the transcription reaction.

    Construct:

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: Resultant viral reporter construct (namely pGL3-DENV2-5′UTR-72ntC-Fluc-3′UTR) and pRL-SV40 vector (Promega), which contains Renilla luciferase gene and serves an internal control reporter construct, were linearized with Xba I. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    Incubation:

    Article Title: Optimized conditions for successful transfection of human endothelial cells with in vitro synthesized and modified mRNA for induction of protein expression
    Article Snippet: At first, 23 μl NTP/cap analog mixture containing 7.5 mM ATP, 1.875 mM GTP (both from MEGAscript® T7 Kit), 7.5 mM Me-CTP, 7.5 mM Pseudo-UTP (both from TriLink BioTechnologies, San Diego, USA), and 2.5 mM 3′-O-Me-m7 G(5′)ppp(5′)G RNA cap structure analog (New England Biolabs, Frankfurt am Main, Germany) was prepared and mixed thoroughly. .. The IVT reaction mixture was incubated at 37°C for 3 h in a thermomixer.

    Article Title: Generation and Functional In Vitro Analysis of Semliki Forest Virus Vectors Encoding TNF-α and IFN-γ
    Article Snippet: The transcribed rRNAs were capped by adding 1 mM 3′-O -Me-m7 G(5′)ppp(5′)G cap-structure analog (Cat. No. S1411S; New England Biolabs, Hitchin, UK) during the transcription reaction. .. The electroporated BHK-21 cells were resuspended in 15 mL of BHK-21 cultivation medium containing 1% FBS (Sigma-Aldrich) and then incubated at 33°C for 48 h. The cell growth medium containing the infectious rSFV particles was then harvested, rapidly frozen in liquid nitrogen, and subsequently used as a source of rSFV particles.

    Luciferase:

    Article Title: Adaptor Protein 1A Facilitates Dengue Virus Replication
    Article Snippet: Translation assay pRL-SV40 vector (Promega), which contains Renilla luciferase gene, was linearized with Xba I. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: Resultant viral reporter construct (namely pGL3-DENV2-5′UTR-72ntC-Fluc-3′UTR) and pRL-SV40 vector (Promega), which contains Renilla luciferase gene and serves an internal control reporter construct, were linearized with Xba I. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    Mass Spectrometry:

    Article Title: Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge
    Article Snippet: Linearized cDNAs were purified via phenol-chloroform extraction, and ∼1 μg was utilized for each transcription reaction: 0.5 mM ribonucleoside triphosphate (rNTP), 0.5 mM m7G(5′)ppp(5′)G RNA cap (NEB), 0.5 μl RNase inhibitor, and 1.25 μl of SP6 polymerase (Ambion, Carlsbad, CA) in a 25-μl total volume. .. Cells were mixed with each transcription mixture (∼10 μg of RNA), placed in 2-mm or 4-mm (Vero cells only) electroporation cuvettes, and immediately electroporated (BTX ECM-830 Square Wave electroporation system; Harvard Apparatus Inc., Holliston, MA) using the following conditions: 680 V, pulse length of 99 μs, interval between pulses of 200 ms, and number of pulses of 5.

    Article Title: Eilat Virus Host Range Restriction Is Present at Multiple Levels of the Virus Life Cycle
    Article Snippet: The transcription reaction mixture contained 0.5 mM ribonucleotide triphosphate (rNTP), 0.5 mM m7G(5′)ppp(5′)G RNA cap (New England BioLabs [NEB]), 0.5 μl RNase inhibitor, and 1.25 μl of SP6 polymerase (Ambion) in a total volume of 25 μl. .. The cells were mixed with each transcription mixture (∼10 μg of RNA), placed in 2-mm or 4-mm (Vero cells only) electroporation cuvettes, and immediately electroporated (BTX ECM-830 Electro Square Porator; Harvard Apparatus Inc.) using the following conditions: 680 V; pulse length, 99 μs; interval between pulses, 200 ms; number of pulses, five.

    Modification:

    Article Title: Dom34 mediates targeting of exogenous RNA in the antiviral OAS/RNase L pathway
    Article Snippet: .. For the synthesis of mRNA whose 5′-terminus was modified with a cap structure, 3′-O-Me-m7G(5′)ppp(5′)G cap analog (New England Biolabs) was added during the T7 polymerase reaction. .. Following DNase I treatment, the synthetic RNAs were purified using a illustra MicroSpin G-25 column (GE Healthcare) or illustra MicroSpin S-400 HR column (GE Healthcare) and quantified using NanoDrop One (Thermo Fisher Scientific).

    Article Title: Combination Immunotherapy of MUC1 mRNA Nano-vaccine and CTLA-4 Blockade Effectively Inhibits Growth of Triple Negative Breast Cancer
    Article Snippet: Nucleotides (ATP and guanosine triphosphate [GTP]) were purchased from Affymetrix, and modified nucleotides (5-methylcytidine-5′-triphosphate and pseudouridine-5′-triphosphate) were purchased from Trilink Biotechnologies. .. 3′-O-Me-m7 G (5′) ppp (5′) G RNA cap structure analog was purchased from NEB.

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: .. IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany). .. Furthermore, 7.5 mM ATP, 1.875 mM GTP (both from MEGAscript T7 Kit), 7.5 mM 5-methylcytidine (5mCTP), 7.5 mM pseudouridine (Ψ) (both from TriLink BioTechnologies, San Diego, CA, USA), and 40 U RiboLock RNase inhibitor (Thermo Scientific, Waltham, MA, USA) were added into the reaction tube.

    Electroporation:

    Article Title: Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge
    Article Snippet: Paragraph title: Transcription and electroporation conditions for virus rescue. ... Linearized cDNAs were purified via phenol-chloroform extraction, and ∼1 μg was utilized for each transcription reaction: 0.5 mM ribonucleoside triphosphate (rNTP), 0.5 mM m7G(5′)ppp(5′)G RNA cap (NEB), 0.5 μl RNase inhibitor, and 1.25 μl of SP6 polymerase (Ambion, Carlsbad, CA) in a 25-μl total volume.

    Article Title: Generation and Functional In Vitro Analysis of Semliki Forest Virus Vectors Encoding TNF-α and IFN-γ
    Article Snippet: RNA Synthesis and Production of Viral Particles RNAs were synthesized and transfected into BHK-21 cells using electroporation as previously described ( ). .. The transcribed rRNAs were capped by adding 1 mM 3′-O -Me-m7 G(5′)ppp(5′)G cap-structure analog (Cat. No. S1411S; New England Biolabs, Hitchin, UK) during the transcription reaction.

    Article Title: Eilat Virus Host Range Restriction Is Present at Multiple Levels of the Virus Life Cycle
    Article Snippet: Paragraph title: Transcription and electroporation conditions for virus rescue. ... The transcription reaction mixture contained 0.5 mM ribonucleotide triphosphate (rNTP), 0.5 mM m7G(5′)ppp(5′)G RNA cap (New England BioLabs [NEB]), 0.5 μl RNase inhibitor, and 1.25 μl of SP6 polymerase (Ambion) in a total volume of 25 μl.

    Transfection:

    Article Title: Adaptor Protein 1A Facilitates Dengue Virus Replication
    Article Snippet: One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. To determine the effect of AP-1A knockdown on translation, Huh7 cells were transfected twice with AP-1A-specific siRNA or control siRNA in a 96-well plate within a 24-h interval using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.

    Article Title: An IGF1R-dependent pathway drives epicardial adipose tissue formation after myocardial injury
    Article Snippet: ModRNAs were in vitro transcribed from plasmid templates (sequences provided in the ) using a custom ribonucleoside blend of 3′-O-Me-m7G(5′)ppp(5′)G cap analog (6 mM, New England Biolabs), guanosine triphosphate (1.5 mM, USB), adenosine triphosphate (7.5 mM, USB), and 5-methylcytidine triphosphate and pseudouridine triphosphate (7.5 mM, TriLink Biotechnologies)as described previously , , RNA was treated with Antarctic Phosphatase (New England Biolabs), quantitated by Nanodrop (Thermo Scientific), precipitated with ethanol and Ammonium Acetate, and resuspended in 10 mM TrisHCl, 1 mM EDTA. .. ModRNA was transfected into cultured cells using RNAiMAX (Life Technologies).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. To determine the effect of hnRNP C1/C2 knockdown on DENV translation, Huh7 cells were transfected twice with hnRNP C1/C2-specific siRNA or control siRNA (286 nM each) in a 96-well plate within a 24-h interval using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer’s instructions.

    Article Title: Generation and Functional In Vitro Analysis of Semliki Forest Virus Vectors Encoding TNF-α and IFN-γ
    Article Snippet: RNA Synthesis and Production of Viral Particles RNAs were synthesized and transfected into BHK-21 cells using electroporation as previously described ( ). .. The transcribed rRNAs were capped by adding 1 mM 3′-O -Me-m7 G(5′)ppp(5′)G cap-structure analog (Cat. No. S1411S; New England Biolabs, Hitchin, UK) during the transcription reaction.

    Cell Culture:

    Article Title: An IGF1R-dependent pathway drives epicardial adipose tissue formation after myocardial injury
    Article Snippet: ModRNAs were in vitro transcribed from plasmid templates (sequences provided in the ) using a custom ribonucleoside blend of 3′-O-Me-m7G(5′)ppp(5′)G cap analog (6 mM, New England Biolabs), guanosine triphosphate (1.5 mM, USB), adenosine triphosphate (7.5 mM, USB), and 5-methylcytidine triphosphate and pseudouridine triphosphate (7.5 mM, TriLink Biotechnologies)as described previously , , RNA was treated with Antarctic Phosphatase (New England Biolabs), quantitated by Nanodrop (Thermo Scientific), precipitated with ethanol and Ammonium Acetate, and resuspended in 10 mM TrisHCl, 1 mM EDTA. .. ModRNA was transfected into cultured cells using RNAiMAX (Life Technologies).

    Generated:

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: Viral translation assay A DENV-2 reporter construct was generated by cloning T7 promoter, DENV-2 5′-UTR, the first 72 nucleotides of DENV-2 capsid-coding sequence, firefly luciferase gene and DENV-2 3′-UTR into pGL3-Basic (Promega) according to a previously described method [ ]. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    Article Title: Optimized conditions for successful transfection of human endothelial cells with in vitro synthesized and modified mRNA for induction of protein expression
    Article Snippet: The generated mRNA transcript is then used to induce protein expression in cells. .. At first, 23 μl NTP/cap analog mixture containing 7.5 mM ATP, 1.875 mM GTP (both from MEGAscript® T7 Kit), 7.5 mM Me-CTP, 7.5 mM Pseudo-UTP (both from TriLink BioTechnologies, San Diego, USA), and 2.5 mM 3′-O-Me-m7 G(5′)ppp(5′)G RNA cap structure analog (New England Biolabs, Frankfurt am Main, Germany) was prepared and mixed thoroughly.

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: PCR products were generated using the following amplification protocol: initial denaturation step at 94°C for 3 min, followed by 25 cycles of denaturation at 94°C for 45 s, annealing at 57°C for 1 min, extension at 72°C for 1 min, and final extension at 72°C for 5 min. .. Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: .. IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany). .. Furthermore, 7.5 mM ATP, 1.875 mM GTP (both from MEGAscript T7 Kit), 7.5 mM 5-methylcytidine (5mCTP), 7.5 mM pseudouridine (Ψ) (both from TriLink BioTechnologies, San Diego, CA, USA), and 40 U RiboLock RNase inhibitor (Thermo Scientific, Waltham, MA, USA) were added into the reaction tube.

    Polymerase Chain Reaction:

    Article Title: Optimized conditions for successful transfection of human endothelial cells with in vitro synthesized and modified mRNA for induction of protein expression
    Article Snippet: In vitro transcription (IVT) After the PCR, the genetic information is in vitro transcribed from DNA to mRNA using MEGAscript® T7 Kit (Life Technologies, Darmstadt, Germany). .. At first, 23 μl NTP/cap analog mixture containing 7.5 mM ATP, 1.875 mM GTP (both from MEGAscript® T7 Kit), 7.5 mM Me-CTP, 7.5 mM Pseudo-UTP (both from TriLink BioTechnologies, San Diego, USA), and 2.5 mM 3′-O-Me-m7 G(5′)ppp(5′)G RNA cap structure analog (New England Biolabs, Frankfurt am Main, Germany) was prepared and mixed thoroughly.

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: Afterward, PCR products were purified using MinElute PCR purification kit (QIAGEN). .. Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: PCR products were then purified using QIAquick PCR Purification Kit (Qiagen, Hilden, Germany) followed by elution in 2 × 10 μL nuclease-free water (Qiagen, Hilden, Germany), and processed for qualitative analysis. .. IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany).

    Recombinant:

    Article Title: Generation and Functional In Vitro Analysis of Semliki Forest Virus Vectors Encoding TNF-α and IFN-γ
    Article Snippet: The linearized plasmids (1 µg of each) were used as templates for the in vitro transcription (SP6 RNA polymerase; Cat. No. AM2071; Thermo Fisher Scientific) of SFV-Helper1 RNA and recombinant RNAs (rRNAs) carrying Ifng, Tnfa , or DsRed . .. The transcribed rRNAs were capped by adding 1 mM 3′-O -Me-m7 G(5′)ppp(5′)G cap-structure analog (Cat. No. S1411S; New England Biolabs, Hitchin, UK) during the transcription reaction.

    Purification:

    Article Title: Adaptor Protein 1A Facilitates Dengue Virus Replication
    Article Snippet: .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. To determine the effect of AP-1A knockdown on translation, Huh7 cells were transfected twice with AP-1A-specific siRNA or control siRNA in a 96-well plate within a 24-h interval using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.

    Article Title: Generation and Characterization of Closely Related Epizootic and Enzootic Infectious cDNA Clones for Studying Interferon Sensitivity and Emergence Mechanisms of Venezuelan Equine Encephalitis Virus
    Article Snippet: Both cDNA clones were purified by using the Qiagen MiniPrep kit. .. RNAs were transcribed in the presence of m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs) by using SP6 RNA polymerase (Invitrogen).

    Article Title: Codon usage regulates protein structure and function by affecting translation elongation speed in Drosophila cells
    Article Snippet: 3′-0-Me-m7G (5′)ppp (5′)G anti-reverse cap structure analog (NEB) was added following the manufacturer's instruction. .. The mRNA products were subsequently purified by LiCl precipitation, quantified by Nanodrop (Thermo Scientific), aliquoted and stored at −80°C before use.

    Article Title: Dom34 mediates targeting of exogenous RNA in the antiviral OAS/RNase L pathway
    Article Snippet: For the synthesis of mRNA whose 5′-terminus was modified with a cap structure, 3′-O-Me-m7G(5′)ppp(5′)G cap analog (New England Biolabs) was added during the T7 polymerase reaction. .. Following DNase I treatment, the synthetic RNAs were purified using a illustra MicroSpin G-25 column (GE Healthcare) or illustra MicroSpin S-400 HR column (GE Healthcare) and quantified using NanoDrop One (Thermo Fisher Scientific).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. To determine the effect of hnRNP C1/C2 knockdown on DENV translation, Huh7 cells were transfected twice with hnRNP C1/C2-specific siRNA or control siRNA (286 nM each) in a 96-well plate within a 24-h interval using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer’s instructions.

    Article Title: Novel Insect-Specific Eilat Virus-Based Chimeric Vaccine Candidates Provide Durable, Mono- and Multivalent, Single-Dose Protection against Lethal Alphavirus Challenge
    Article Snippet: .. Linearized cDNAs were purified via phenol-chloroform extraction, and ∼1 μg was utilized for each transcription reaction: 0.5 mM ribonucleoside triphosphate (rNTP), 0.5 mM m7G(5′)ppp(5′)G RNA cap (NEB), 0.5 μl RNase inhibitor, and 1.25 μl of SP6 polymerase (Ambion, Carlsbad, CA) in a 25-μl total volume. ..

    Article Title: eIF4A alleviates the translational repression mediated by classical secondary structures more than by G-quadruplexes
    Article Snippet: This kit uses the anti-reverse cap analogue (ARCA) to ensure the cap is in the correct orientation. mRNA was purified using the MEGAclear kit (ThermoFisher AM1908) and quantified by nanodrop. .. For , 7.5 mM ATP/CTP/UTP, 1.5 mM GTP and either 6 mM ARCA (NEB S1411S) or 6 mM G(5′)ppp(5′)A RNA Cap Structure Analog (NEB S1406S) was used whereas for Figure , 7.5 mM ATP/CTP/UTP, 6 mM ARCA (S1411S) and either 1.5 mM GTP or 1.5 mM 7-deazaguanine (TriLink N-1044) was used.

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: Afterward, PCR products were purified using MinElute PCR purification kit (QIAGEN). .. Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Eilat Virus Host Range Restriction Is Present at Multiple Levels of the Virus Life Cycle
    Article Snippet: Linearized cDNAs were purified via phenol-chloroform extraction, and ∼1 μg was utilized for each transcription reaction. .. The transcription reaction mixture contained 0.5 mM ribonucleotide triphosphate (rNTP), 0.5 mM m7G(5′)ppp(5′)G RNA cap (New England BioLabs [NEB]), 0.5 μl RNase inhibitor, and 1.25 μl of SP6 polymerase (Ambion) in a total volume of 25 μl.

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: PCR products were then purified using QIAquick PCR Purification Kit (Qiagen, Hilden, Germany) followed by elution in 2 × 10 μL nuclease-free water (Qiagen, Hilden, Germany), and processed for qualitative analysis. .. IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany).

    Sequencing:

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: Viral translation assay A DENV-2 reporter construct was generated by cloning T7 promoter, DENV-2 5′-UTR, the first 72 nucleotides of DENV-2 capsid-coding sequence, firefly luciferase gene and DENV-2 3′-UTR into pGL3-Basic (Promega) according to a previously described method [ ]. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    De-Phosphorylation Assay:

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany). .. Subsequently, dephosphorylation was performed using the Antarctic Phosphatase Kit (New England Biolabs, Frankfurt am Main, Germany) to remove 5′-triphosphates, and the synthetic mRNA was purified again using the RNeasy Kit in 2 × 40 μL nuclease-free water.

    Plasmid Preparation:

    Article Title: Adaptor Protein 1A Facilitates Dengue Virus Replication
    Article Snippet: Translation assay pRL-SV40 vector (Promega), which contains Renilla luciferase gene, was linearized with Xba I. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    Article Title: An IGF1R-dependent pathway drives epicardial adipose tissue formation after myocardial injury
    Article Snippet: .. ModRNAs were in vitro transcribed from plasmid templates (sequences provided in the ) using a custom ribonucleoside blend of 3′-O-Me-m7G(5′)ppp(5′)G cap analog (6 mM, New England Biolabs), guanosine triphosphate (1.5 mM, USB), adenosine triphosphate (7.5 mM, USB), and 5-methylcytidine triphosphate and pseudouridine triphosphate (7.5 mM, TriLink Biotechnologies)as described previously , , RNA was treated with Antarctic Phosphatase (New England Biolabs), quantitated by Nanodrop (Thermo Scientific), precipitated with ethanol and Ammonium Acetate, and resuspended in 10 mM TrisHCl, 1 mM EDTA. .. ModRNA was transfected into cultured cells using RNAiMAX (Life Technologies).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: Resultant viral reporter construct (namely pGL3-DENV2-5′UTR-72ntC-Fluc-3′UTR) and pRL-SV40 vector (Promega), which contains Renilla luciferase gene and serves an internal control reporter construct, were linearized with Xba I. .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany).

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: To synthesize the mRNA, first of all, DNA templates were generated by PCR using 50–100 ng plasmid DNA, 0.7 μM forward primer (5′-TTGGACCCTCGTACAGAAGCTAATACG-3′) and 0.7 μM reverse primer (5′-T120 CTTCCTACTCAGGCTTTATTCAAAGACCA-3′) (Ella Biotech, Martinsried, Germany), and HotStar HiFidelity Polymerase Kit (QIAGEN, Hilden, Germany), according to the manufacturer’s instructions. .. Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Unbiased screen of RNA tailing activities reveals a poly(UG) polymerase
    Article Snippet: pCS2 3HA MS2-MUT-2 (MAP136) was linearized with Sac II, and 3 μg of linearized plasmid was transcribed with Ampliscribe SP6 High Yield Transcription Kit (Epicentre), according to manufacturer’s instructions. pLGMS2-luc (RNA with three MS2-binding sites) , was linearized with Bgl II, and 1 μg of linearized plasmid was transcribed with T7 Flash In Vitro Transcription Kit (Epicentre), according to manufacturer’s instructions. .. Transcription reactions included m7 G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs).

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: Briefly, pcDNA 3.3 plasmid (Aldevron, Fargo, ND, USA) insert encoding eGFP and pEX-K4 plasmid insert encoding Link N (Eurofins Genomics GmbH, Ebersberg, Germany) were amplified using the Hot Star HiFidelity Polymerase Kit (Qiagen, Hilden, Germany), according to manufacturer’s instructions. .. IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany).

    Agarose Gel Electrophoresis:

    Article Title: Generation and Characterization of Closely Related Epizootic and Enzootic Infectious cDNA Clones for Studying Interferon Sensitivity and Emergence Mechanisms of Venezuelan Equine Encephalitis Virus
    Article Snippet: RNAs were transcribed in the presence of m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs) by using SP6 RNA polymerase (Invitrogen). .. The yield and integrity of the RNA transcripts were monitored by agarose gel electrophoresis.

    Article Title: Codon usage regulates protein structure and function by affecting translation elongation speed in Drosophila cells
    Article Snippet: 3′-0-Me-m7G (5′)ppp (5′)G anti-reverse cap structure analog (NEB) was added following the manufacturer's instruction. .. Denaturing agarose gel electrophoresis was used to determine the quality of the mRNA products.

    Article Title: eIF4A alleviates the translational repression mediated by classical secondary structures more than by G-quadruplexes
    Article Snippet: Integrity of RNA was checked by inspection on a native agarose gel. mRNAs used in and Figure were transcribed using the TranscriptAid T7 High Yield Transcription Kit (ThermoFisher K0441) as per manufacturer's instructions. .. For , 7.5 mM ATP/CTP/UTP, 1.5 mM GTP and either 6 mM ARCA (NEB S1411S) or 6 mM G(5′)ppp(5′)A RNA Cap Structure Analog (NEB S1406S) was used whereas for Figure , 7.5 mM ATP/CTP/UTP, 6 mM ARCA (S1411S) and either 1.5 mM GTP or 1.5 mM 7-deazaguanine (TriLink N-1044) was used.

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany). .. The purity and specific length of generated DNA templates and mRNA products were analyzed using 1% agarose gel electrophoresis at 100 V for 45 min and staining with 1× GelRed (Biotium, Fremont, CA, USA) in 1× Tris-borate-EDTA (TBE) buffer.

    In Vitro:

    Article Title: Adaptor Protein 1A Facilitates Dengue Virus Replication
    Article Snippet: .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. To determine the effect of AP-1A knockdown on translation, Huh7 cells were transfected twice with AP-1A-specific siRNA or control siRNA in a 96-well plate within a 24-h interval using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.

    Article Title: Codon usage regulates protein structure and function by affecting translation elongation speed in Drosophila cells
    Article Snippet: Paragraph title: In vitro transcription assay ... 3′-0-Me-m7G (5′)ppp (5′)G anti-reverse cap structure analog (NEB) was added following the manufacturer's instruction.

    Article Title: Dom34 mediates targeting of exogenous RNA in the antiviral OAS/RNase L pathway
    Article Snippet: Paragraph title: In vitro mRNA synthesis ... For the synthesis of mRNA whose 5′-terminus was modified with a cap structure, 3′-O-Me-m7G(5′)ppp(5′)G cap analog (New England Biolabs) was added during the T7 polymerase reaction.

    Article Title: An IGF1R-dependent pathway drives epicardial adipose tissue formation after myocardial injury
    Article Snippet: .. ModRNAs were in vitro transcribed from plasmid templates (sequences provided in the ) using a custom ribonucleoside blend of 3′-O-Me-m7G(5′)ppp(5′)G cap analog (6 mM, New England Biolabs), guanosine triphosphate (1.5 mM, USB), adenosine triphosphate (7.5 mM, USB), and 5-methylcytidine triphosphate and pseudouridine triphosphate (7.5 mM, TriLink Biotechnologies)as described previously , , RNA was treated with Antarctic Phosphatase (New England Biolabs), quantitated by Nanodrop (Thermo Scientific), precipitated with ethanol and Ammonium Acetate, and resuspended in 10 mM TrisHCl, 1 mM EDTA. .. ModRNA was transfected into cultured cells using RNAiMAX (Life Technologies).

    Article Title: Role of human heterogeneous nuclear ribonucleoprotein C1/C2 in dengue virus replication
    Article Snippet: .. One μg of purified DNA was subjected to in vitro transcription using the RiboMAX Large Scale RNA Production System-T7 (Promega) in the presence of 20 mM m7 G(5′)ppp(5′)G RNA cap structure analog (New England BioLabs, Ipswich, MA, USA) and resultant RNA product was purified using RNeasy Mini Kit (QIAGEN, Hilden, Germany). .. To determine the effect of hnRNP C1/C2 knockdown on DENV translation, Huh7 cells were transfected twice with hnRNP C1/C2-specific siRNA or control siRNA (286 nM each) in a 96-well plate within a 24-h interval using Lipofectamine RNAiMAX (Invitrogen) according to the manufacturer’s instructions.

    Article Title: Optimized conditions for successful transfection of human endothelial cells with in vitro synthesized and modified mRNA for induction of protein expression
    Article Snippet: Paragraph title: In vitro transcription (IVT) ... At first, 23 μl NTP/cap analog mixture containing 7.5 mM ATP, 1.875 mM GTP (both from MEGAscript® T7 Kit), 7.5 mM Me-CTP, 7.5 mM Pseudo-UTP (both from TriLink BioTechnologies, San Diego, USA), and 2.5 mM 3′-O-Me-m7 G(5′)ppp(5′)G RNA cap structure analog (New England Biolabs, Frankfurt am Main, Germany) was prepared and mixed thoroughly.

    Article Title: Generation and Functional In Vitro Analysis of Semliki Forest Virus Vectors Encoding TNF-α and IFN-γ
    Article Snippet: The linearized plasmids (1 µg of each) were used as templates for the in vitro transcription (SP6 RNA polymerase; Cat. No. AM2071; Thermo Fisher Scientific) of SFV-Helper1 RNA and recombinant RNAs (rRNAs) carrying Ifng, Tnfa , or DsRed . .. The transcribed rRNAs were capped by adding 1 mM 3′-O -Me-m7 G(5′)ppp(5′)G cap-structure analog (Cat. No. S1411S; New England Biolabs, Hitchin, UK) during the transcription reaction.

    Article Title: eIF4A alleviates the translational repression mediated by classical secondary structures more than by G-quadruplexes
    Article Snippet: Paragraph title: In vitro transcription ... For , 7.5 mM ATP/CTP/UTP, 1.5 mM GTP and either 6 mM ARCA (NEB S1411S) or 6 mM G(5′)ppp(5′)A RNA Cap Structure Analog (NEB S1406S) was used whereas for Figure , 7.5 mM ATP/CTP/UTP, 6 mM ARCA (S1411S) and either 1.5 mM GTP or 1.5 mM 7-deazaguanine (TriLink N-1044) was used.

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: Paragraph title: In Vitro mRNA Synthesis ... Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany).

    Article Title: Unbiased screen of RNA tailing activities reveals a poly(UG) polymerase
    Article Snippet: Paragraph title: In vitro Transcription ... Transcription reactions included m7 G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs).

    Article Title: Exogenous Delivery of Link N mRNA into Chondrocytes and MSCs—The Potential Role in Increasing Anabolic Response
    Article Snippet: Subsequently, in vitro transcription (IVT) was performed to generate mRNA from the DNA product. .. IVT was conducted using MEGAscript T7 Kit (Ambion, Glasgow, Scotland) and the generated mRNA was modified using 2.5 mM 3’-0-Me-m7G(5’)ppp(5’)G RNA Cap Structure Analog (New England Biolabs, Frankfurt am Main, Germany).

    Produced:

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: Next, the mRNA was produced by in vitro transcription (IVT) using MEGAscript T7 Kit (Ambion, Glasgow, Scotland), according to the manufacturer’s instructions. .. Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany).

    Concentration Assay:

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany). .. After each reaction step, the mRNA was purified using RNeasy kit (QIAGEN), and the mRNA concentration was adjusted to 100 ng/μL in nuclease-free water.

    Staining:

    Article Title: Improving the Angiogenic Potential of EPCs via Engineering with Synthetic Modified mRNAs
    Article Snippet: Furthermore, 2.5 mM 3′-0-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (New England Biolabs, Frankfurt, Germany) was used for 5′ end capping, and the mRNA was dephosphorylated using 5 U/mL Antarctic phosphatase (New England Biolabs, Frankfurt am Main, Germany). .. The purity and specific length of generated DNA templates and mRNA products were analyzed using 1% agarose gel electrophoresis at 100 V for 45 min and staining with 1× GelRed (Biotium, Fremont, CA, USA) in 1× Tris-borate-EDTA (TBE) buffer.

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    New England Biolabs 3 o me m7g
    3 O Me M7g, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 95/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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