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Applications and Protocols

Armored Technology™: Applications and Protocols

The main advantage of Armored RNA controls is that the packaged RNA is stable and ribonuclease-resistant in plasma and other matrices. The RNA is compatible with any RNA-based clinical assay after sample extraction.

Applications
Armored RNA® Armored RNA®
Quant™
Assay Development Laboratories lack access to all viruses X X
Daily Controls Positive controls to verify assay performance X X
Internal Controls Normalize data X X
Calibrators Determine mean range of assay X
Quantitate Quantitate against standard curve X
Extraction Control Determine extraction efficiency X
X

Direct Detection of Target
External RT-PCR Control
  • Heat lyse Armored RNA or Armored RNA Quant at 75°C for 3 minutes
  • Place on ice
  • Detect by RT-PCR
    • Use 5 µL of heat-lysed Armored RNA or desired amount of Armored RNA Quant for a 20 µL RT reaction
    • Use 2 to 5 µL of RT for a 25 to 50 µL PCR reaction
Sample Quantitation – Standard Curve
  • Spike a known quantity of Armored RNA Quant into negative matrix
  • Prepare standard curve by 10-fold serial dilutions (at least 5 points)
  • Extract standard curve along with patient samples via kit of choice
  • Perform 3-5 replicates
  • Detect by qRT-PCR
  • Quantitate samples against standard curve

Specimen Extraction Efficiency
Internal Control (IC)
Sequence of IC different from that of target
  • Spike a known quantity of Armored RNA or Armored RNA Quant into patient specimen
  • Extract sample via commercial kit of choice
  • Detect IC by RT-PCR
External Extraction Control
  • Spike either Armored RNA or Armored RNA Quant into negative matrix (e.g. serum, plasma)
  • Extract sample via kit of choice
  • Detect target by RT-PCR