TRANSCRIPTME RNA Kit (RT31)

- Overview
  - The TRANSCRIPTME RNA Kit is a system which includes all the necessary components for synthesizing first-strand cDNA, apart from the template total RNA or mRNA. The synthesized single-stranded cDNA is suitable for real-time quantitative RT-PCR applications. The TRANSCRIPTME RNA Kit has been formulated to provide high yields of full-length cDNA product and to increase sensitivity in RT-qPCR. Starting material can range from 10 pg to 5 μg of total RNA.
    
    The kit includes both the TRANSCRIPTME Enzyme Mix, with a recombinant thermostable reverse transcriptase M-MuLV without RNase H activity and the RIBOPROTECT RNase Inhibitor. The optimal reaction buffer with a combination of random hexamers and oligo(dT)18 primers, MgCl2 and dNTPs mix is also provided for increased sensitivity.
    
    TRANSCRIPTME Reverse Transcriptase has increased thermal stability, which allows the reaction to be carried out at a higher temperature (optimum activity at 50°C). It increases the efficiency and specificity of those transcribed RNA regions which are rich in GC pairs and/or contain secondary structures. The enzyme has no 3’→5’ exonuclease or RNase H
    activity, which improves synthesis of a full-length cDNA, even from long mRNA templates, using random priming. The enzyme gives high yields of first strand cDNA up to 10 kb long.
    
    Use of the recombinant RNase inhibitor RIBOPROTECT protects the RNA template from degradation. The enzyme selectively hydrolyzes the phosphodiester bonds of RNA only when it is hybridized to DNA. The RNase H does not degrade either single and doublestranded DNA or unhybridized RNA. This optional step can improve the sensitivity of subsequent RT-qPCR reaction since the PCR primers will bind more easily to the cDNA.
    Using of the RNase H is recommended only when it contributes to a full-length cDNA synthesis and increased yields of first-strand cDNA.
    
    Features and advantages
    • High yields of full-length cDNA products (up to 10 kb)
    • Formulated to increase sensitivity in RT-qPCR and RT-PCR assays
    • Fewer pipetting steps – minimized contamination risk
    • Simultaneous cDNA synthesis with mRNA and rRNA templates
    • Starting material: 10 pg – 5 μg of total RNA or 10 pg – 500 ng of mRNA
    • Optimal reaction temperature: 50°C
    • The reverse transcriptase shows no RNase H or 3’→5’ exonuclease activity, has supreme thermostability and is suitable for difficult RNA templates
    • RNase inhibitor protects the RNA template from degradation
    • Additional RNase H treatment may increase the sensitivity of the RT-qPCR reaction
    
    Please contact us at for specific academic pricing.
- Properties
  - Storage
    
    Store all components at -20°C. The stability of the reagents can be extended if stored at -70°C. The product is stable until the expiry date providing it is stored and used correctly.
    
    Shipping
    
    Shipping on dry or blue ice.
- Applications
  - Application
    
    cDNA synthesis; cDNA library construction; RNA analysis
    
    Application Description
    
    The TRANSCRIPTME RNA Kit has been formulated to provide high yields of full-length cDNA product and to increase sensitivity in RT-qPCR. Starting material can range from 10 pg to 5 μg of total RNA.; optimal reaction temp. 50°C; contains Enzyme Mix (M-MuLV reverse transcriptase and RNase inhibitor); 2x Master Mix (oligodT primers, random hexamers, dNTPs, MgCl2) and RNase H in a separate tube (optional step)
- Reference
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    Vitamin C, A and E supplementation decreases the expression of HSPA1A and HSPB1 genes in the leukocytes of young polish figure skaters during a 10-day training camp