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    Friday, 01 March 2013 13:06

    AccuPower RocketScript RT PreMix

    Rate this item
    (1 Vote)

    AccuPower RocketScript RT PreMix contains Gentaur’s exclusive M-MLV based thermostable reverse transcriptase (RTase), RocketScript. Native M-MLV RTase has maximum activity at relatively low temperatures (42°C), causing several problems in reverse transcription of RNA molecules with complex secondary structure. 

    RocketScript has thermostable activity (42°C~70°C), allowing efficient cDNA synthesis from virtually any RNA. The lyophilized PreMix contains all components necessary for a successful reverse transcription reaction, including RTase, RNase inhibitor and buffer components. Just add template RNA, primers and water, and the RT reaction is ready to go.

    accupower-gentaur-antibodies-aproketscript overview

     

    Schematic representation of the 5’UTR of a gene, with complex secondary structure, at three different temperatures. Note that RocketScript shows full activity at 70°C allowing it to synthesize the complete gene sequence where M-MLV and other Reverse Transcriptase's fail. 

    Features and Benefits 

    Thermostable Activity: RocketScript is able to perform reverse transcription reactions throughout a wide range 
    of temperatures from 42°C to 70°C. 

    Enhanced Performance: RocketScript has enhanced performance to handle both high and low input RNA 
    concentrations as well as short and long RT target sizes. 

    Easy-to-use: The product contains the enzyme itself, plus RNase inhibitors and all other components necessary for the best reverse transcription results in the tube. 

    Reproducibility: Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment. 

    Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided. 

    Stability: Stable at room temperature for a month one year at 4°C and for 2 years in a -20°C freezer 

    RNase, DNase and Proteinase-free: Ensures the integrity of your samples. 

    Broad range of working temperatures: For RNAs that are G:C rich or significant secondary structure 

    Stable for 2 years at -20°C: Long shelf life 

    Specifications

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


    Application

    - First-strand synthesis of cDNA from RNA molecules (RT)
    - RT-PCR
    - Random priming reaction
    - Library construction
    - Probe labeling
    - mRNA 5end Mapping by Primer Extension Analysis
    - Real time PCR

    accupower-gentaur-antibodies-aproketscript figure1

    Figure 1. Sensitivity comparison between AccuPower RocketScript RT PreMix and M-MLV RTase
    Sensitivity results of AccuPower RocketScript RT PreMix using GAPDH compared with conventional Reverse transcriptases. 
    Each 100 ng – 10 fg of total RNA used for RT and the same amount of RT products used for electrophoresis.
    Lane 1: 10 fg Human total RNA from HeLa cell           Lane 2: 100 fg Human total RNA from HeLa cell
    Lane 3: 1 pg Human total RNA from HeLa cell            Lane 4: 10 pg Human total RNA from HeLa cell
    Lane 5: 100 pg Human total RNA from HeLa cell        Lane 6: 1 ng Human total RNA from HeLa cell
    Lane 7: 10 ng Human total RNA from HeLa cell          Lane 8: 100 ng Human total RNA from HeLa cell

    accupower-gentaur-antibodies-aproketscript figure2

    Figure 2. Comparison of amplification efficiency between AccuPower RocketScript RT PreMix and competitorsM-MLV RTase.
    RocketScript is able to handle a wide range of sample concentrations and transcript lengths so your downstream applications are minimally effected by the reverse transcription step.
    Lanes 1-3: 1,000 ng, 100 ng and 10 ng of total RNA from HeLa cells, respectively.

    accupower-gentaur-antibodies-aproketscript figure3

    Figure 3. Sensitivity comparison between AccuPower RocketScript RT PreMix and competitor RTases using Real Time PCR
    Reverse transcription conditions: conventional 1 hr incubation at 60°C, deactivation at 95°C for 5 min All cDNAs were amplified with AccuPower DualStar™ qPCR PreMix (K-6110) from Gentaur.

    Concentration RocketScript RT PreMix Supplier Q Supplier I
    10,000 23.91 25.63 24.43
    1,000 27.33 28.92 28.03
    100 30.62 32.42 30.88
    10 33.63 35.43 33.95
    Efficiency 104% 103% 108%
    Linearity 0.99999 0.9996 0.9995

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