qPCR Extraction Control Orange

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 assay, but also to validate the success of the extraction step, reducing the chance of obtaining a false negative result in the sample DNA.

Category:qPCR & RT-qPCR Extraction Controls Brand: Bioline

Description

Product Highlights

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. 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.

Applications

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

Specifications

Components

Reagent	2000 Reactions
Internal Control DNA Orange*	20 x 500 µL
25x Control Mix 560(containing Cal Fluor® Orange 560 labeled probe)	20 x 100 µL

* The Internal Control DNA is in viable E. coli cells (genotype: F- deoR endA1 recA1 relA1 gyrA96 hsdR17(r_k-, mk+) supE44 thi-1 phoA Δ(lacZYA-argF)U169Ф80lacZΔ15λ–pBR322 (ranseqb1 AmpR)).

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.