DNA extraction from S. pneumoniae isolates was carried out by alkaline lysis as described previously [16 (link)]. For DNA extraction from whole blood, initial pretreatment of the samples was performed as described elsewhere [17 (link)]. Briefly, 100 μL of whole-blood EDTA was added to 100 μL of Tris-EDTA buffer containing 75 U/mL of mutanolysin (Sigma Chemical Co., St. Louis, MO), and the mixture was incubated for 1 hour at 37°C. DNA extraction was then carried out with the DNeasy 96 Blood & Tissue Kit (Qiagen, Venlo, the Netherlands), according to the manufacturer’s instructions.
Protocol detail
DNA Extraction from S. pneumoniae and Whole Blood
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Corresponding Organization :
Other organizations : Armauer Hansen Research Institute, Ghent University, Addis Ababa University, St. Paul's Hospital Millennium Medical College
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Variable analysis
independent variables
- DNA extraction method (alkaline lysis for S. pneumoniae, pretreatment and DNeasy 96 Blood & Tissue Kit for whole blood)
- DNA yield/quality (not explicitly mentioned)
- Incubation time (1 hour at 37°C)
- Mutanolysin concentration (75 U/mL)
- Positive control: Not mentioned
- Negative control: Not mentioned
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