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.
| Component | 50 rxn kits | Storage |
| 5× Reaction Buffer* | 200μL | -20℃ |
| 100mM ATP Solution | 100μL | -20℃ |
| 100mM GTP Solution | 100μL | -20℃ |
| 100mM UTP Solution | 100μL | -20℃ |
| 100mM CTP Solution | 100μL | -20℃ |
| Enzyme Mix | 75μL | -20℃ |
| Control Template(0.5μg/μL) | 10μL | -20℃ |
| DNase I (RNase-free, 1U/μL) | 50μL | -20℃ |
| Nuclease-free H2O | 1mL | -20℃ |
| Ammonium Acetate Stop Solution | 1.5mL | -20℃ |
| Lithium Chloride Precipitation Solution | 1.5mL | -20℃ |
| Gel Loading Buffer | 0.5mL | -20℃ |
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Note: 5× 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.
| Component | Amount |
| Nuclease-free H2O | to 20μL |
| 100mM ATP Solution | 2μL |
| 100mM GTP Solution | 2μL |
| 100mM CTP solution | 2μL |
| 100mM UTP solution | 2μL |
| Template DNA | 1μg* |
| 5× Reaction Buffer | 4μL |
| Enzynme Mix | 1.5μL |
*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 >150μg 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.