EZ1 Virus Mini Kit v2.0 (48)

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Publication protocol

Parallel harmonization testing of the newly implemented automated EZ1 nucleic acid extraction system: From the same samples, nucleic acids were extracted and enriched using the automated EZ1 system (Qiagen). EZ1-based extraction from the frozen EDTA-blood samples with the EZ1 DNA Blood 200 µl Kit (Qiagen) was performed exactly as described by the manufacturer.
For stool samples, pretreatment with ASL-buffer (Qiagen) and InhibitEx tablets (Qiagen) to reduce inhibiting effects of the matrix was performed as described by the manufacturer for the QIAamp DNA Stool Mini Kit (Qiagen). In detail, ASL-buffer was preheated at 70 °C to dissolve precipitates. Afterwards, either 200–300 mg formed stool or 200 µl of unformed stool were vortex-mixed with 1.4 ml ASL-buffer for 1 min. Then, the samples were again heated at 70 °C for 5 min, followed by vortex-mixing for 15 s and centrifugation at 20.000g for 1 min. A total of 1.2 ml supernatant was transferred to another laboratory cup, and the pellet was discarded. The InhibitEx tablet was added and vortex-mixing was performed for 1 min until it was completely dissolved. Afterwards, the samples were incubated at room temperature for 1 additional minute before they were centrifuged at 20.000g for 6 min. The supernatant was transferred into another laboratory cup, and the pellet was discarded. Again, the samples were centrifuged at 20.000g for 3 min, before 200 µl of the supernatants was used for nucleic acid extraction with the EZ1 Virus Mini Kit v2.0 (Qiagen) as described by the manufacturer.

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