For intraoperative NDMA analysis, the tumor biopsy was immediately placed in lysis buffer (Qiagen) at the operating room, allowing digestion to begin during transport to the laboratory.10 (link) Upon arrival, the biopsy lysate was homogenized for 30 s (TissueLyser, Qiagen) and digested for 10 min at 56°C and 5 min at 70°C. This reduced the total sample handling and DNA extraction time to 30 min. The DNA quantity and quality were still above the required standards.
Rapid DNA Extraction for NDMA Analysis
For intraoperative NDMA analysis, the tumor biopsy was immediately placed in lysis buffer (Qiagen) at the operating room, allowing digestion to begin during transport to the laboratory.10 (link) Upon arrival, the biopsy lysate was homogenized for 30 s (TissueLyser, Qiagen) and digested for 10 min at 56°C and 5 min at 70°C. This reduced the total sample handling and DNA extraction time to 30 min. The DNA quantity and quality were still above the required standards.
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Corresponding Organization : Berlin Institute of Health at Charité - Universitätsmedizin Berlin
Other organizations : Humboldt-Universität zu Berlin, Freie Universität Berlin, University of Latvia
Variable analysis
- Extraction method using spin columns (QIAamp DNA Micro Kit, Qiagen, NL)
- DNA quantity (measured using Qubit 4.0 fluorometer)
- DNA purity (assessed using NanoDrop 260/280 ratio)
- Tissue sample size (~15 mg of tumor tissue)
- Lysis buffer (Qiagen) used for immediate digestion of tumor biopsy
- Homogenization time (30 s)
- Digestion time (10 min at 56°C and 5 min at 70°C)
- Total sample handling and DNA extraction time (30 min)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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