VLP-RNA Extraction Control Monitor the extraction, reverse-transcription and amplification with a single control that can be customized to work with your assay primers
Meridian’s VLP-RNA Extraction Controls offer a simple and effective way to reduce false negative results in RT-qPCR assays. The biomimetic controls are composed of virus-like particle (VLP) shells packaged with a defined number of copies of target RNA molecules. Their composition closely resembles a test sample and serves as a full-process control, monitoring the success the of an RT-qPCR assay from lysis/extraction to reverse transcription and amplification. The VLP can be customized with specific RNA sequences of your choice, up to a total of 1000nt, to work with your assay primers and enable multiple controls within a single VLP. Or alternatively, Meridian offers a universal, ready-to-use control (and matching primers) which contains an RNA sequence with no known homology. The VLP controls are compatible with commonly used RNA extraction methods and they can be lyophilized to remove cold-chain requirements for room-temperature stable kits.
VLP-RNA Extraction Control
• Contains a defined number of copies of target RNA molecules, encapsidated within a virus-like particle (VLP) • RNA sequence is customizable up to 1000nt • Closely mimics the test sample, undergoing the same processing from lysis and extraction to RT-qPCR detection • Non-infectious material for ease of handling and shipping • Compatible with commonly used RNA extraction methods and lyophilization for creating freeze-dried mixes
1 mL (~1x10 4 copies/µL) 20 mL (~1x10 4 copies/µL) 1 mL (~1x10 4 copies/µL) 20 mL (~1x10 4 copies/µL)
VLP-RNA Extraction Control Red
VLP-RNA Extraction Control Orange
VLP-RNA Extraction Control MDX071
POSSIBLE CAUSES OF NEGATIVE DETECTION IN A RT-qPCR ASSAY.
Sample is not lysed, DNA/RNA is not released from the sample
MDx assay not functional or inhibited, DNA/RNA signal not detected
No Signal No detection
Target DNA/RNA absent from the sample
Extraction not functional, DNA/RNA is not isolated
DNA/RNA is degraded
Recommended lysis conditions
Monitors quality of sample lysis step Amplification traces of VLP-RNA Extraction Control isolated using the ISOLATE II RNA Mini Kit with the recommended (Red) or sub-optimal (Green) lysis conditions. The RT-qPCR results demonstrate how the VLP-RNA Control Extraction can help to monitor the quality of the sample lysis step.
Sub-optimal lysis conditions
Maintains stability in the presence of nucleases found in biofluids VLP-RNA Extraction Controls were lyophilized and incubated at ambient temperature for 24 hours (dark blue); incubated at ambient temperature for 24 hours (light blue) or spiked directly into the lysis buffer (grey) for blood, plasma and serum samples. The RT-qPCR results illustrate that the VLP-RNA Extraction Control protects packaged RNA from degradation by nucleases in these biofluids until they are ready for analysis.
Lyophilized VLP incubated 24h at room temp with biofluid | VLP incubated 24h at room temp with biofluid | VLP spiked directly in the lysis buffer
Highly stable and lyophilization compatible
B) Thermal resistance stability. VLP-RNA Extraction Controls were pre-treated at different temperatures for 10 minutes and then analyzed.
A) Stable after freezing/ thawing cycles. VLP-RNA Extraction Controls were submitted to various freeze/thaw cycles: 0 (blue), 1 cycle (black), 5 cycles (red), 10 cycles (grey) and 20 cycles (green) and then analysed. C) Shelf-life. VLP-RNA Extraction Controls were stored for a month, in different conditions, ambient temperature (black), 4°C (green) and -20°C (red) before being analyzed.
D) Lyophilization compatible. VLP-RNA Extraction Control are resistant to nucleases even when lyophilized. Lyophilized and nuclease treated (blue), not lyophylized and nuclease treated,(orange) not lyophilzed and no nuclease treated (red).
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