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Larova is a German original manufacturer of PCR reagents including dNTPs from milli-litre ... multi-litre scale.
This issue: Macromolecular Contaminants
The most critical macromolecules potentially present in dNTPs are human DNA and bacterial DNA: Enzymes of bacterial origin are commonly used during enzymatic dNTP synthesis, and human DNA is ubiquitously present during dNTP handling. Since the presence of only a few copies of such genomic DNA may result in false positive PCR, we analyse each dNTP lot at the very end of the production process using a highly sensitive qPCR assay (Figure 1). In addition, testing for residual enzymatic activities (DNases, RNases, Nicking) is performed with FRET-probes, and a colorimetric UV-assay is used for ensuring absence of protease activity.
Figure 1: A multiplex qPCR assay verifies absence of contaminating bacterial or human DNA. Bacterial DNA is amplified using primers and probes for the 16S rRNA gene. Human DNA is detected by amplification of a beta-actin gene fragment. Traces of contaminating DNA are typically detected at ct-values in between 35 and 45.
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| Responsible for content / Imprint | ||||||
LAROVA GmbH
Rheinstr. 17B 14513 Teltow Germany Tel: +49 – 3328 – 3956 0 Fax: +49 – 3328 – 3956 39 E-Mail: info@larova.com Register Court: Amtsgericht Potsdam, HRB 17 114 P VAT No.: DE 231 513 961 Managing Director: Dr. Mathias Grün |
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| www.larova.com | ||||||
