【RC1000】EzRNA™ RNA Capping System, 50 RXN

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Description

The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5' end of RNA to form m7Gppp5'N-RNA (Cap-0 RNA). The 2'-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2' -OH of the first nucleotide at the 5' end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.


Features

  • 2’-O-Methyltransferase included for Cap-1 RNA

  • High capping efficiency

  • High stability

  • RNase inhibitor is included to enhance the stability of capping reaction


Application

  • Generation of 5’Cap-0 (m7Gppp) and Cap-1 (m7GpppNm-) RNA by enzymatic reaction

  • mRNA synthesis for in vitro translation

  • Gene expression studies

  • mRNA vaccine development and therapeutics


Storage

-20°C for 24 months

Highly expression of eGFP-mRNA in mammalian cells

Sample 1 for three columns


The eGFP-mRNA was generated utilizing the SMOBIO EzRNATM T7 High Yield RNA Synthesis Kit (Ψ-UTP) in conjunction with the EzRNATM RNA Capping System. Subsequently, eGFP-mRNA was transfected into HeLa cells using Lipofectamine 3000 for an incubation period of 18 hours. Higher than 90% transfection efficiency was observed.

Contents

Components

Volume

Vaccinia Capping Enzymes

50 μl

2´-O-Methyltransferase

50 μl

10X Capping Buffer

100 μl

S-adenosylmethionine (SAM) (32 mM)

50 μl

RNase Inhibitor (20 U/μl)

50 μl

GTP (10 mM)

50 μl

Nuclease-Free Water

1 ml


Storage

-20°C for 12 months




  1.  For prevention of RNase contamination, it is highly advised to wear gloves, utilize nuclease-free tubes and reagents, and meticulously clean pipettes and bench surfaces throughout the procedure.

  2. Prior to the capping reaction, it is crucial to guarantee the purification of RNA, which should be suspended in nuclease-free water devoid of EDTA and salts.

  3. Subjecting the RNA to heating at 65°C for 5 minutes before initiating the capping reaction eliminates secondary structures present at the 5´ end of the transcript. Increase the duration to 10 minutes for RNA known to have highly structured at 5’ ends.

  4. Due to its instability at pH 7–8, 37°C, please dilute SAM immediately before initiating the reaction.

Odoo - Sample 1 for three columns

[IT1000] EzRNA™T7 High Yield RNA Synthesis Kit, 50 RXN

Application

  • Generation of RNA from T7 promoter-driven DNA sequences

  • Suitable for subsequent cap-0 and cap-1 modification


Odoo - Sample 1 for three columns

[IT1100] EzRNA™T7 High Yield RNA Synthesis Kit (Ψ-UTP), 50 RXN

Application

  • Generation of RNA from T7 promoter-driven DNA sequences

  • Suitable for subsequent cap-0 and cap-1 modification

Odoo - Sample 1 for three columns

[IT1200] EzRNA™ T7 High Yield RNA Synthesis Kit (me1Ψ-UTP), 50 RXN

Application

  • Generation of RNA from T7 promoter-driven DNA sequences

  • Suitable for subsequent cap-0 and cap-1 modification


 

Odoo - Sample 1 for three columns

[RI1000] RNAok™ RNase Inhibitor

Application

  • RT-PCR

  • cDNA Synthesis 

  • in vitro transcription

 

Odoo - Sample 3 for three columns

FluoroVue™ Nucleic Acid Gel Stain

  • Excellent for in-gel staining

  • Sensitivity up to 0.14 ng (DNA) or or 1 ng (total RNA)

  • A safe alternative to EtBr

  • Suitable for blue or UV light