Checkout View Cart Items
Total RNA Purification

SuperScript® VILO™ cDNA Synthesis Kit

http://products.invitrogen.com/ivgn/en/US/adirect/invitrogen?cmd=catProductDetail&productID=11754050
The SuperScript® VILO™ cDNA Synthesis Kit contains tried and true SuperScript® III Reverse Transcriptase (RT) formulated in an enhanced buffer system containing RNaseOUT™ Recombinant Ribonuclease Inhibitor and a proprietary helper protein. You'll get the most reliable first-strand synthesis and higher cDNA yields.

Excellent linearity

  • Looking for consistent results for your RT reactions? Unprecedented linearity  across the broadest range of input material?  You'll get them with the SuperScript® VILO™ cDNA Synthesis Kit:
    • For probe-based two-step qRT-PCR, linear cDNA yields from 1pg to 2.5ug of total RNA in 20ul reactions
    • For SYBR® GreenER™ Two-Step qRT-PCR, linear cDNA yields from 1pg to 100ng of total RNA in 20ul reactions
    Reaction volumes can be scaled up to 100ul if more cDNA is needed, while maintaining the same linearity of the reaction.

    Excellent linearity in your RT step means you will:
    • obtain the same relative representation in your cDNA qPCR template, regardless of gene abundance
    • during normalization, get both your gene of interest and the reference gene in a linear phase for cDNA synthesis even if their abundances differ in the starting material
    • get greater accuracy in your qRT-PCR data by reducing this common bias
    .



Figure 3. The SuperScript® VILO™ cDNA Synthesis Kit provides reliable performance across an extended linear range of RNA input. Serial dilutions of total RNA from HeLa cells were reverse transcribed using the SuperScript® VILO™ cDNA Synthesis Kit, followed by triplicate qPCR reactions using human β-actin TaqMan® assays (Applied Biosystems) with the EXPRESS qPCR SuperMix Universal (Invitrogen) on an ABI PRISM® 7900HT real-time PCR system. The standard curve (A) shows that even outside the recommended input of 1 pg to 2.5 μg, the kit exhibits a coefficient of correlation of 0.996 for up to 5 μg of input RNA. The amplification plot (B) illustrates that the triplicates are aligned across all dilutions, demonstrating the robustness of the SuperScript® VILO™ cDNA Synthesis Kit over a broad linear range of RNA input. Normalize your lower-abundance genes to your reference genes without worrying about potential variation of RT efficiency at different RNA input levels.

Increased yield and reduced qPCR inhibition

  • The SuperScript® VILO™ cDNA Synthesis Kit generates far greater yields of cDNA, coupled with less downstream qPCR inhibition. This means you can:
    • archive cDNA for future studies
    • use greater amounts of cDNA in qPCR to increase sensitivity up to 4-fold without fear of inhibiting the reaction

    This benefit has been achieved through a careful balancing of SuperScript® III RT, RNaseOUT™ RNase inhibitor, and the addition of a proprietary helper protein, providing a clear advantage in your qRT-PCR reagents if you have a low-copy target.
  • Figure 4. The SuperScript® VILO™ cDNA Synthesis Kit demonstrates greater yields than competitors. Total RNA (2 μg and 1 ng) from HeLa cells was reverse transcribed using either the SuperScript® VILO™ cDNA Synthesis Kit (blue) or Mulv competitor kit (green), followed by triplicate qPCR reactions using β-actin–specific TaqMan® assays (Applied Biosystems) with the EXPRESS qPCR SuperMix Universal (Invitrogen) on an ABI PRISM® 7900HT real-time PCR system. At both high and low RNA input levels, the SuperScript® VILO™ cDNA Synthesis Kit outperformed the Mulv competitor kit by at least 2 cycles, indicating a 4-fold increase in sensitivity.

Detection flexibility

  • Get the most sensitive qPCR performance with the SuperScript® VILO™ cDNA Synthesis Kit regardless of detection method. Using either probe of SYBR® based real-time PCR detection, can expect up to 8-fold sensitivity increases, compared to other available kits. SuperScript® VILO™ cDNA Synthesis Kits are available separately, or as part of our new EXPRESS line of qPCR and qRT-PCR reagents designed for fast cycling, real-time PCR instruments.



Figure 5. The SuperScript® VILO™ cDNA Synthesis Kit can be successfully used with SYBR® Green– or fluorogenic probe/primer–based qPCR detection chemistries. A. Equal amounts (10 pg) of total RNA from HeLa cells were reverse transcribed using Invitrogen’s SuperScript® VILO™ cDNA Synthesis Kit (blue), Bio-Rad’s iScript™ cDNA Synthesis Kit (yellow), Qiagen’s QuantiTect® Reverse Transcription Kit (green), and Stratagene’s AffinityScript® QPCR cDNA Synthesis Kit (pink). The resulting cDNA was then amplified using hsp70 gene–specific primers in triplicate using the Invitrogen SYBR® GreenER™ qPCR SuperMix on an ABI PRISM® 7900HT real-time PCR system. The SuperScript® VILO™ cDNA Synthesis Kit exhibits higher sensitivity than the other kits when used with SYBR® Green detection, as shown by the Cts that are obtained at least 2 cycles earlier. B. Equal amounts (1 μg) of total RNA from HeLa cells were reverse transcribed using the same kits named in A. The cDNA obtained was then amplified in triplicate TfR gene–specific TaqMan® assays (Applied Biosystems) using the EXPRESS qPCR SuperMix with Premixed ROX on an ABI PRISM® 7900HT real-time PCR system. The SuperScript® VILO™ cDNA Synthesis Kit exhibits higher sensitivity than the other kits when used with probe-based detection, as shown by the Cts that are obtained at least 2 cycles earlier.