- Simple - streamlined protocol for straightforward validation of DNA extraction and determination of qPCR assay inhibition
- Sensitive - control assay identifies even small effects on DNA extraction and inhibition of amplification
- Optimized - control DNA has a sequence with no known homology to any organism thereby avoiding detection of sample DNA
- Specific - probe-based assay designed specifically for real-time PCR assays
- Flexible - ideal for use with a wide range of sample types, including inhibitor-rich samples like blood, urine and sputum samples
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. Bioline has developed the DNA Extraction Control (DEC), which more closely mimics the test sample, as compared to spike controls. Genetic material from the test sample and the DEC is simultaneously extracted by common extraction methods, with the extraction control being as sensitive to inhibition and extraction failure as the test sample.
The DEC 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 DEC 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. DEC also monitors co-purification of PCR inhibitors that may cause biased or false amplification patterns.
- Pathogen detection
- Cancer risk assessment
- Gene expression analysis
- Copy number variation (CNV) analysis
- Viral loading
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DNA Extraction Control 670 is suitable for use with commercially available silica-membrane DNA extraction kits and CHELEX matrices and has been tested on a wide range of qPCR platforms including ABI-7500, LightCycler 480®, RotorGene-Q™, Mic and MX3005P®.
DNA Extraction Control 670 uses Quasar®670 and is also available with Cal Fluor® Orange 560 or Cal Fluor® Red 610, to fit in with existing protocols. CAL Fluor and Quasar dyes are performance-optimized fluorophores for multiplex qPCR.
Certification of Analysis (COAs)
Internal Control DNA*
5 x 500 µL
20 x 500 µL
25x Control Mix (containing Quasar® 670 labeled probe)
5 x 100 µL
20 x 100 µL
* The Internal Control DNA is in viable α-Select E. coli cells (genotype: F- deoR endA1 recA1 relA1 gyrA96 hsdR17(rk-, mk+) supE44 thi-1 phoA Δ(lacZYA-argF)U169Ф80lacZΔ15λ–pBR322 (ranseqb1 AmpR)).
Storage & Stability
All kit components should be stored at -20°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.
Shipped on dry/blue ice.