QIAamp 96 PowerFecal QIAcube HT Kit (5)

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Publication protocol

Bacterial DNA extraction: DNA was extracted from 200-μL frozen MBRA suspension using a QIAamp 96 PowerFecal QIAcube HT kit from Qiagen Laboratories (Venlo, Netherlands) with mechanical disruption (bead-beating). Briefly, 650 μL of prewarmed buffer PW1 was added to 50 μL of each sample. Samples were thoroughly homogenized using bead-beating with a TissueLyser before centrifuging the plate at 4000 rpm for 5 min at 20 °C in order to pellet beads and particles. Four hundred microliters of supernatant was added into a new 96-well plate containing 150 μL of Buffer C3. After mixing and incubation on ice for 5 min, centrifugation was performed at 4000 rpm for 5 min at 20 °C. Three hundred microliters of each supernatant was added to a new 96-well plate, and 20 μL of Proteinase K was added and incubated for 10 min at room temperature. The following steps were next performed on a QIAcube high-throughput robot: addition of 500 μL of Buffer C4, DNA binding to a QIAamp 96 plate, column wash using AW1 (800 μL), AW2 (600 μL), and ethanol (400 μL), and DNA elution using ATE buffer (100 μL).

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Download the product protocol from Qiagen for QIAamp 96 PowerFecal QIAcube HT Kit (5) below.

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