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High Yield T7 RNA Synthesis Kit

  • Cat.No.ON-040
  • SourceN/A
  • PurityN/A
  • Storage-20℃
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Product description: The High Yield T7 RNA Synthesis Kit is designed to produce 25-fold more full-length RNA transcript per reaction than conventional in vitro transcription reactions. The yield is up to 210μg RNA per reaction.

Materials provided with the kit:

The kit contains sufficient reagents for 50 reaction of 20μL each.

Component50 rxn kitsStorage
10× Reaction Buffer*100μL-20℃
100mM ATP Solution100μL-20℃
100mM GTP Solution100μL-20℃
100mM UTP Solution100μL-20℃
100mM CTP Solution100μL-20℃
Enzyme Mix100μL-20℃
Control Template(0.5μg/μL)10μL-20℃
DNase I (RNase-free, 1U/μL)50μL-20℃
Nuclease-free H2O1mL-20℃
Ammonium Acetate Stop Solution1.5mL-20℃
Lithium Chloride Precipitation Solution1.5mL-20℃
Gel Loading Buffer0.5mL-20℃

Related Products:

Poly(A) Polymerase, Cat#ON-126

Vaccinia Capping Enzyme, Cat#ON-028

mRNA Cap-2'-O-Methyltransferase, Cat#ON-014

RNase III, E.coli, Cat#ON-024

Note:10× Reaction Buffer store at -70℃ may result in the formation of a white precipitate. If this happens, heat the tube to 37℃ for 5 minutes and mix thoroughly to resuspend the precipitate.

1. The following amounts are for a single 20 μL reaction. Reactions may be scaled up or down if desired. 

Nuclease-free H2Oto 20μL
100mM ATP Solution2μL
100mM GTP Solution2μL
100mM CTP solution2μL
100mM UTP solution2μL
Template DNA1μg*
10× Reaction Buffer2μL
Enzynme Mix2μL

*The transcript may vary depending on the amount of template DNA.

2. Mix thoroughly and incubate at 37℃ 2h.

3. (optional)Add 1μL DNase I (RNase-free), mix well and incubate 30 min at 37°C.

Storage Conditions: -20℃

Quality Assurance: All components are tested in a functional assay as described in this procedure. A 20μL reaction containing 1 μg of the control template DNA which codes for a ~800b transcript synthesized >150 μg of RNA after a 2 hr incubation.

1. Milligan JF, Groebe DR, Witherell GW, and Uhlenbeck OC (1987) Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA template. Nucl. Acids Res. 15: 8783–8798.

2. Molecular Cloning, A Laboratory Manual, 2nd edition. (1989) editor C Nolan, Cold Spring Harbor Laboratory Press.

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