RocketScript™ Reverse Transcriptase

Features and Benefits

• Thermostable Activity: Native M-MLV reverse transcriptase has low thermostable activity, therefore restricting reverse transcription reactions to relatively low temperatures (42°C). This attribute prevents RNA molecules containing many stems and loops (complex secondary structures) from being efficiently transcribed. To resolve this shortcoming, Bioneer has utilized synthetic biotechnology to develop a RTase that is active even at high temperatures of 60°C and above. By removing the traditional reaction temperature limit of 42°C, you are now able to choose your reaction temperature from 42°C –70°C and optimize your cDNA synthesis experiments.

• Enhanced Performance: While engineering the enzyme, the researchers at Bioneer have gone ahead and engineered robust performance into RocketScript. You can now confidently perform experiments with target lengths long and short, or input RNA concentrations low and high knowing that if the RNA is in the sample, it will be accurately represented in the cDNA.

• Ease-of-use: All components necessary for cDNA synthesis including thermostable RTase and RNase inhibitor are included in the product for ease-of-use. All you need is source RNA and primers, and preparation for your reverse transcription reaction is complete.

• Reproducibility: Each batch is produced under strict quality controls. Errors that commonly occur during mass production are eliminated during the individual packaging process. Bioneer’s current batch processing system allows for the production of more accurate and reproducible end-product yield.

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment Size: Up to 10 kb

Application

Gene synthesis
- First-strand synthesis of cDNA from RNA molecules (Reverse Transcription)
- RT-PCR
- Random priming reactions
- Library construction
- Probe labeling
- mRNA 5’-end mapping by primer extension analysis
- Real time PCR

Experimental data


Figure 1. Increasing reverse transcription reaction temperature increases yield of long targets.
TFRC (full-length: 5.2 kb) was reverse transcribed with A: Normal M-MLV; and B: RocketScript at 1) 100 ng and 2) 10 ng total RNA concentrations complex RNA amplification


Figure 2. RocketScript retains full activity up to 70 °C.
A low copy transcript (Myc: 495 bp) was reverse transcribed at the indicated temperatures. Thermostability of RocketScript Reverse Transcriptase is outstanding compared to a leading competitor’s reverse transcriptase. Lanes 1-4 are 100 ng, 10 ng, 1 ng and 0.1 ng of total RNA from HeLa cells, respectively


Figure 3. Comparison of amplification quality between RocketScript RTase and competitor RTases Sensitivity test. Target gene expression Level.

Lane M: 1 kb DNA Ladder
Lane 1: 100 ng Human total RNA from HeLa cell
Lane 2: 10 ng Human total RNA from HeLa cell
Lane 3: 1 ng Human total RNA from HeLa cell
Lane 4: 100 pg Human total RNA from HeLa cell

• RocketScript Reverse Transcriptase

MSDS

• RocketScript Rtase

• RocketScript Rtase-dilution buffer

• RocketScript RTase-reaction buffer

• 100mM DTT

Brochure

• RocketScript Reverse Transcriptase - 2011 Brochure

Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001:2008 - certificate

• EN ISO 13485:2003 / AC:2009 - certificate