Formerly DNA Extraction Control 560. DNA 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.
ISOLATE II RNA Mini kit was used to extract the RNA from HeLa cells containing A) spiked control DNA and B/ REC. To show inefficient extraction, the lysis buffer and/or binding buffer substituted with PBS. The results demonstrate complete lysis (yellow line), no lysis buffer (brown), no binding buffer (grey), no lysis and no binding buffer (pink). The results illustrate that the spiked control DNA is insensitive to extraction failure, whereas REC is sensitive, with the Ct values being affected.
ISOLATE II RNA Mini kit was used to extract the RNA from HeLa cells containing REC. Increasing concentrations of EDTA were included in the reaction to simulate increasing levels of inhibition. The results illustrate that REC is increasingly inhibited by increasing concentrations of EDTA, showing that inhibition of PCR reaction can be identified using REC.
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 Orange* 25x Control Mix 560 |
1 x 200 µL
20 x 100 µL |
Storage & Stability |
All components should be stored at -80°C upon receipt. When stored under the recommended conditions and handled correctly, full activity is retained until the expiry date on the outer box label. |
| Shipping Conditions | Shipped on dry ice. |
Cat. No. Size
MDX027 2000 Reactions
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