Contamination Control

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PCR Quantification Standards

The quantification standard is genomic DNA extracted from low passage and defined mycoplasma or legionella cultivated in liquid medium according to the European Pharmacopoeia or in Hayflick´s medium. The extraction of the DNA is done by column absorption methods and the DNA purified by phenol/chloroform extraction and ethanol precipitation. The concentration was determined photometrically and by qPCR. The DNA concentration was adjusted by dilution with DNA stabilizing buffer. Genomic DNA is used as a control template when DNA amplification tests by conventional PCR are performed (e.g. as contamination control in cell culturing). With the qPCR technology, this titrated DNA standard with a defined concentration of genome copies is used as calibrator to generate standard curves. Therefore the provided solution is diluted in a 10x dilution series with Tris-HCI buffer and used as a PCR sample. With the generated fluorescent data, the software provided with the qPCR instrument calculates a standard curve, which can be used to determine the DNA concentration of unknown samples.

Applications

These DNA-Standards allow for the performance of a control reaction for PCR using qPCR instrumentation and to create a standard curve for quantification experiments, i.e. with Venor^®Mp Mycoplasma pneumoniae Diagnostic Kit for qPCR. The DNA standards can be used as a general control to confirm PCR system performance using a qPCR instrument or for the validation of detection limits in respect to conventional PCR tests.

Contents

1 vial (green cap) of Standard DNA, 100 µl, contains approx. 10⁶ genomes/µl.
3 vials (white cap) with 1 ml of DNA free water.

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