Takara NucleoSpin cfDNA Midi \48 Preps

NucleoSpin cfDNA Midi is designed for the rapid, medium-scale preparation of highly pure cell-free DNA from up to 5 mL of blood plasma using vacuum filtration. The purification protocol involves lysis of 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 DNA to the silica membrane, impurities are removed by several washing steps and cfDNA is eluted after a final drying step. The use of vacuum filtration technology eliminates tedious centrifugation steps and manual handling of columns during processing. NucleoSpin cfDNA Midi allows for processing of up to 24 samples in parallel in 90 min.

Optimized protocol for cfDNA isolation from Cell-Free DNA BCT (Streck) tubes.Blood was drawn from five subjects and preserved with Cell-Free DNA BCT (Streck) and EDTA tubes, respectively. cfDNA was isolated with NucleoSpin cfDNA Midi. cfDNA yields from EDTA and Streck tubes were similar due to optimization of the kit protocol for processing samples preserved in Streck tubes. DNA yields were quantified by qPCR.

Efficient isolation of cfDNA from 2 mL of human EDTA-treated plasma.cfDNA was isolated from individual EDTA plasma samples with the NucleoSpin cfDNA Midi kit and a vacuum-based kit from a competitor (Competitor Q). DNA yields were quantified by qPCR.

NucleoSpin Plasma Midi procedure: binding and washing step

NucleoSpin Plasma Midi procedure: drying step

NucleoSpin Plasma Midi procedure: elution step