qPCR Extraction Control Red

This product replaces former catalog numbers BIO-35028-BL and BIO-35029-BL

DNA Extraction Control not only enables users of a diagnostic qPCR assay to determine, if there are inhibitors in the PCR assay, but also to validate the success of the extraction step, reducing the chance of obtaining a false negative result in the sample DNA.

• 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 - DNA Extraction Control consists of viable Alpha Select E. coli cells of a known concentration, containing plasmid pBR322 with the Internal Control DNA sequence, which has no homology to any organism
• 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

Product Description

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.

Applications

• Pathogen detection
• Cancer risk assessment
• Gene expression analysis
• Copy number variation (CNV) analysis
• Genotyping
• Viral loading

Illustration of the extraction process

DEC assesses effects of extraction as well as inhibition throughout the entire workflow.