VLP-RNA Extraction Controls
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Why use VLP-RNA Extraction Controls in an RT-qPCR assay?
Possible causes of negative detection in an RT-qPCR
Spike-In internal controls are RNA molecules added between the nucleic acid isolation and the detection, they do not monitor the extraction and lysis steps. Mimics internal controls are RNA molecules provided in a format resembling a biological sample (i.e. micelles) and are added just before lysis, but do not allow long incubation with the sample. VLP-RNA Extraction Control have better nuclease resistance, for long incubation with the sample, better quantification for consistent input in MDx processes and are easier to ship and store, ensuring that the MDx process provides reliable results by questioning each step of the process from lysis, through extraction to detection.
-Contains a defined number of copies of target RNA molecules, encapsidated within a non-infectious virus-like particle (VLP).
-Closely mimics the test sample, undergoing the same processing from lysis and extraction to RT-qPCR detection.
-Compatible with commonly used RNA extraction methods and lyophilization for creating freeze-dried mixes.
Description
VLP-RNA Extraction Control can be customized with specific RNA sequences of your choice, up to a total of 1,000 nt, to work with your assay primers and probe to enable multiple controls within a single VLP. The detection of VLP-RNA Extraction Control acts as a PCR positive control and confirms the success of the extraction, reverse transcription, and amplification steps, avoiding the misinterpretation of a false negative result.
Specifications
| Description | An Internal Control RNA sequence of the customers choice, encapsidated in a virus-like particle, used to confirms the success of the extraction, reverse transcription, and amplification steps |
| Application | one-step RT-qPCR |
| Sample type | Tissue, cells |
| Presentation | 1 vial |
| Storage | -20 °C |
| Mix stability | See outer label |
VLP-RNA Extraction Controls contain an internal control RNA sequence, with no known homology to any organism, encapsidated in a virus-like particle. The VLP-RNA Extraction Control is spiked in the sample prior to RNA extraction. Following RNA extraction, VLP-RNA Extraction Control can be detected in RT-qPCR by adding the VLP Detection Mix (containing specific primers and probe (emission wavelength = 560nm or 670nm)) to the reaction mix. The detection of VLP-RNA Extraction Control acts as a PCR positive control and confirms the success of the extraction, reverse transcription, and amplification steps, avoiding the misinterpretation of a false negative result.
Catalogs & Brochures
Reagent Solutions for Molecular Diagnostics Reagent Solutions for Molecular Diagnostics
VLP-RNA Extraction Control VLP-RNA Extraction Control
VLP Extraction Control Custom Project VLP Extraction Control Custom Project
FAQs: VLP-RNA Extraction Control
The VLP-RNA Extraction Controls are in plant based VLPs, so there is no animal/human viruses used within the process, so there is no possibility of the VLP-RNA Extraction Control giving false positive results.
The VLP-RNA Extraction Controls are extremely robust and stable quantitative PCR positive control, we have tested then for 20 cycles of freeze thawing, storage at ambient temperature for one month, heating to 50 ᵒC for 10 minutes, lyophilization and nuclease treatment without seeing any change in Ct values.
The primers and probe for the VLP-RNA Extraction Control are in the VLP Detection Mix, however, we do not supply the primers and probe for the custom VLP-RNA Extraction Control.
The two colors are for the fluorescent dyes on the VLP-RNA Extraction Control probe. VLP Detection Mix Red (Cy5 – emission wavelength = 670nm) and VLP Detection Mix Orange (HEX – emission wavelength = 555nm). This allows you to choose one that will fit with your existing protocol in a multiplex RT-qPCR assay.
No, the VLP-RNA Extraction Control will work with commercially available silica-membrane extraction kits and CHELEX matrices and has been tested on a wide range of qPCR platforms.
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