Formerly DNA Extraction Control 670. qPCR Extraction Control not only enables users of a diagnostic qPCR assay to determine if there are inhibitors in the PCR
REC assesses effects of extraction as well as inhibition throughout the entire workflow.
A/ β 2 Microglobulin (b2MG) was amplified in triplicate from human genomic DNA in singleplex (green) and in duplex with Internal Control (blue). B/ The Internal Control was amplified in singleplex (red) and in duplex with b2MG (blue). The Cts show no difference between singleplex (b2MG – green, Internal Control – red) and duplex (blue) reaction assays in both target gene and Internal Control.
A/ A fragment of the β2MG gene was amplified from genomic DNA and B/ the internal control sequence from the DEC. Increasing concentrations of EDTA were included in the reaction to simulate increasing concentrations of an inhibitor. The results illustrate that DEC gives the same pattern of inhibition as with the sample target, showing that inhibition of PCR reaction can be identified using DEC.
A common practice in qPCR is to add a known amount of spiked control DNA after DNA extraction, this monitors PCR inhibition but has no value as an extraction control. The ideal situation is to have the test sample and internal control undergo the same processing prior to qPCR. Meridian has developed the qPCR Extraction Control, which more closely mimics the test sample, as compared to spike controls. Genetic material from the test sample and the qPCR Extraction Control is simultaneously extracted by common extraction methods, with the extraction control being as sensitive to inhibition and extraction failure as the test sample.
The qPCR Extraction Control cells are of a known concentration, containing the Internal Control DNA sequence. This sequence contains no known homology to any organism and, importantly, has minimal interference with detection of sample DNA. The qPCR Extraction Control cells are spiked into the lysis buffer with the target sample, prior to DNA extraction. Control Mix, which includes primers and probe, is then added to the reaction mix before amplification. Signal derived from the Internal Control DNA confirms the success of the extraction step. qPCR Extraction Control also monitors co-purification of PCR inhibitors that may cause biased or false amplification patterns.
| Reagent | 2000 Reactions |
| Internal Control DNA Red* 25x Control Mix 670 |
20 x 500 µL 20 x 100 µL |
| Storage & Stability | All kit components should be stored at -80°C upon receipt. When stored under the recommended conditions and handled correctly, full activity of the kit is retained until the expiry date on the outer box label. |
| Shipping Conditions | Shipped on dry ice. |
Cat. No. Size
MDX026 2000 Reactions
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