BK MM Strain Human Polyomavirus Quantitated Viral DNA

Virus: BK Human Polyomavirus

Virus Preparation: Gel purified linear viral DNA from plasmid containing entire BKV genome. The BK viral genome is linearized at the BamH I site (base 4856) in the VP 1 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)

PCR Analysis of DNA Control: PCR analysis was performed on purified BK polyomavirus DNA using the PE Biosystems GeneAmp^® Gold PCR Reagent Kit and primers specific to the large T antigen of BK polyomavirus. The reaction produced a 474 bp fragment.

DNA Quantitation: Copy number of quantitative viral DNA controls is determined by real time PCR using the Roche LightCycler^®. DNA copy number may vary dependent 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 quantitative DNA controls, 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, quantitative DNA controls are not intended for use as negative controls for other organisms.