JCV MAD1 Strain Quantitated Viral DNA

Virus: JC Human Polyomavirus

Virus Preparation: Gel purified linear viral DNA from plasmid containing entire JCV genome. The JC viral genome is linearized at the BamH I site (base 4307) in the large T antigen gene. Note: Due to the linear nature of this DNA, primer sets that span the BamH I digestion site will not produce an amplification product.

Suspending Buffer: 10 mM Tris, 1 mM EDTA, pH 8.0 with 50 µg/mL glycogen carrier.

DNA Copy Number: 1-2 × 10⁵ copies/µL (varies by lot, please contact ABI for details)

CAUTION: ABI does not recommend storage of dilutions of these controls under any conditions. All dilutions should be made immediately before use and used promptly. We have observed that dilutions used for standard curves may tend to “lose” copy number with time (sometimes a matter of an hour or so after dilution), especially at dilutions less than 100-1000 copies per microliter.

PCR Analysis of DNA Control: PCR analysis was performed on purified JC Polyomavirus DNA using the PE Biosystems GeneAmp^® Gold PCR Reagent Kit and primers specific to the large T antigen of JC Polyomavirus. The reaction produced a 215 bp fragment.

DNA Quantitation: Viral DNA copy number is determined by real time PCR using the Roche LightCycler^®. DNA copy number may vary depending on the quantitation method used.

Application: ABI’s quantitated DNA controls are prepared from virus, bacteria, parasites, or mollicutes, and are intended for use as positive PCR quantitation standards for the organism in question.  Due to the nature of these products, ABI cannot guarantee their suitability as extraction controls.  Additionally, due to the extreme sensitivity of detection in PCR reaction, and since no method of purification can guarantee the complete absence of extraneous agents, DNA controls are not intended for use as negative controls for other organisms.