The purification protocol is based on the lysis of the plasma samples using Proteinase K incubation in the presence of a chaotropic salt-containing buffer. Following the initial lysis step, a binding buffer is added to the samples and the lysate is loaded onto the binding columns. After loading and vacuum filtration-driven binding of the DNA to the silica membrane, impurities are removed by several washing steps, and cfDNA is eluted after a final drying step (Figure 1).
NucleoSpin cfDNA Midi
Isolation of cell-free DNA from up to 5 ml of blood plasma
- Superior recovery of fragmented cell-free DNA
- Parallel purification of 24 samples—special adapter set for NucleoVac 96 Vacuum Manifold
- Optimized protocol for Cell-Free DNA BCT (“Streck Tubes”)
NucleoSpin cfDNA Midi is designed for rapid preparation of highly pure cell-free DNA from up to 5 ml of blood plasma using vacuum filtration. The application of vacuum filtration technology eliminates tedious centrifugation steps and manual handling of columns during the process (e.g., disposal of collected column flowthrough). Combining NucleoSpin cfDNA Midi with the NucleoVac 96 Vacuum Manifold and the Starter Set Midi enables processing of up to 24 samples in parallel in ~90 minutes.
Efficient isolation of cfDNA from plasma
The NucleoSpin cfDNA Midi kit was used to isolate cfDNA from 2 ml of human EDTA plasma in comparison with a competitor kit (Figure 2).
Optimized protocol for Cell-Free DNA BCT (“Streck Tubes”)
The NucleoSpin cfDNA Midi kit provided similar yields of cfDNA from blood samples preserved with Cell-Free DNA BCT (“Streck Tubes”) and EDTA tubes, due to optimization of the protocol for the “Streck Tubes” (Figure 3).
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