• Linear quantitation of mRNA targets over 10⁶-fold range of input RNA
  
  
• Quantitate transcripts from as little as 500 fg total RNA – 1/20th of a single cell!
  
  
• Fast, flexible RT step complete in 15 min; Use reaction temperatures up to 50ºC
  
  
• Compatible with TaqMan®, SYBR® Green, and gel-based detection methods
  
  

Optimizing RT Temperature for qRT-PCR All RNA templates are not created equal. Some contain regions of highly structured RNA that can impede the processivity of RT enzymes during reverse transcription. Increasing the reaction temperature in these cases can often “melt out” secondary structure and expedite readthrough by the RT. However, some reverse transcriptases lose significant activity when the reaction temperature is increased above 42ºC. The MessageSensor RT, in contrast, maintains superb target sensitivity at temperatures of up to 50ºC, thereby allowing higher reaction temperatures that can help reduce secondary structure (see lower bar chart at left).

MessageSensor: RNA to PCR Results in 2 hrs
MessageSensor is designed to provide unsurpassed results in the least amount of time. The kit contains a unique formulation of MMLV RT developed for a single tube approach to create a highly optimized protocol and reagent system. The result is a kit that delivers rapid results without compromising lower-end sensitivity. Simply prepare a master mix that combines your primers (and dye-based reporter) with your RNA sample, DNA polymerase, the MessageSensor RNase H⁺ reverse transcriptase, buffer and nucleotides, and you are ready to go. The reverse transcription step takes only 15 minutes — you can go from RNA sample to PCR results in less than 2 hours. Although the protocol is short, MessageSensor won’t compromise your limit of detection: human GAPDH mRNA can be readily detected from as little as 500 fg (see figure at right) of total RNA— or about 1/20th of the total RNA in a single cell.

Linear Detection Over a Million-fold Range of Input RNA
Effective qRT-PCR requires the reliable detection of targets over a broad range of input RNA amounts. The figure at right demonstrates that the amplification reaction produces linear results (R²=0.998; triplicate samples) across a million-fold range of inputs - 500 fg to 0.5 μg of total RNA, when human GAPDH mRNA was quantified using MessageSensor. Unlike other kits that require different enzymes for low and high amounts of input RNA, MessageSensor provides a one-size-fits-all solution for your most demanding qRT-PCR applications. Real-time or endpoint detection and quantitation of mRNA targets can be performed on poly(A) RNA, total RNA, or synthetic transcripts.

A Complete RT Kit for qRT-PCR Applications
Unlike many other qRT-PCR kits, MessageSensor includes a total RNA control, a control human GAPDH primer set, RNase inhibitor, and nucleotides, as well as a buffer additive that enables detection with SYBR® Green dye. The MessageSensor Kit is provided with enough reagents for 50 reactions. When purchased in combination with Ambion’s SuperTaq™ Real-Time, the MessageSensor RT Kit provides all the components you need for one-step qRT-PCR (except for gene-specific primers and fluorogenic probe or dye) in a single convenient order.

A detailed instruction manual with separate protocols for performing one-step RT-PCR using TaqMan® probes, SYBR® Green and end-point detection is provided with the kit.

Trademarks, Patents, Licensing