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Product overview

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.

Protocol overview Application data Product-at-a-glance

Protocol overview  

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 Plasma Midi procedure: drying step
NucleoSpin Plasma Midi procedure: binding and washing step
NucleoSpin Plasma Midi procedure: elution step

Figure 1. The NucleoSpin cfDNA Midi procedure.

Application data  

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).

Efficient isolation of cfDNA from 2 mL human EDTA plasma

Figure 2. Efficient isolation of cfDNA from 2 ml of human EDTA 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 (Quantifiler Human DNA Quantification Kit, Thermo Fisher Scientific on an Applied Biosystems 7500 Real-Time PCR System, Thermo Fisher Scientific). For each sample processed, NucleoSpin cfDNA Midi provided DNA yields that were either greater than or comparable to the yields obtained with the kit from Competitor Q.

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).

Optimized cfDNA isolation protocol from Streck Cell-Free DNA BCT tubes

Figure 3. Optimized protocol for cfDNA isolation from Cell-Free DNA BCT (“Streck Tubes”). Blood was drawn from five independent test subjects and preserved using both Cell-Free DNA BCT (“Streck Tubes”) and EDTA tubes. cfDNA was isolated from samples preserved in both tube types using the NucleoSpin cfDNA Midi kit. cfDNA yields from EDTA tubes and “Streck Tubes” were similar. DNA yields were quantified by qPCR (Quantifiler Human DNA Quantification Kit, Thermo Fisher Scientific on an Applied Biosystems 7500 Real-Time PCR System, Thermo Fisher Scientific).

Product-at-a-glance  

Technology Silica membrane
Format Midi spin columns
Starting material 1–5 ml plasma
Blood draw tubes EDTA, Cell-Free DNA BCT (Streck)
Fragment size ≥50 bp
Typical yield Varies depending on sample source, storage, and quality
Elution volume 200 µl (140 µl final eluate volume)
Preparation time 90 min/24 preps (EDTA plasma)

Related products

Cat. # Product Size License Quantity Details
740303.48 NucleoSpin® cfDNA Midi 48 Preps USD $1010.00

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.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents You May Also Like Image Data

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Optimized protocol for cfDNA isolation from Cell-Free DNA BCT (Streck) tubes

Optimized protocol for cfDNA isolation from Cell-Free DNA BCT (Streck) tubes

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.

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Efficient isolation of cfDNA from 2 ml of human EDTA-treated plasma

Efficient isolation of cfDNA from 2 ml of human EDTA-treated plasma

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.

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NucleoSpin Plasma Midi procedure: binding and washing step

NucleoSpin Plasma Midi procedure: binding and washing step

NucleoSpin Plasma Midi procedure: binding and washing step

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NucleoSpin Plasma Midi procedure: drying step

NucleoSpin Plasma Midi procedure: drying step

NucleoSpin Plasma Midi procedure: drying step

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NucleoSpin Plasma Midi procedure: elution step

NucleoSpin Plasma Midi procedure: elution step

NucleoSpin Plasma Midi procedure: elution step

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740303.48: NucleoSpin DNA Plasma Midi

740303.48: NucleoSpin DNA Plasma Midi
740302.48 NucleoSpin® cfDNA Midi Core Kit 48 Preps USD $830.00

The NucleoSpin cfDNA Midi Core Kit includes all of the components provided with the NucleoSpin cfDNA Midi kit (Cat. # 740303.48) minus the 24-Square-well Block 10 mL, and 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. The NucleoSpin cfDNA Midi Core Kit allows for processing of up to 24 samples in parallel in 90 min.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents You May Also Like Image Data

Back

Optimized protocol for cfDNA isolation from Cell-Free DNA BCT (Streck) tubes

Optimized protocol for cfDNA isolation from Cell-Free DNA BCT (Streck) tubes

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.

Back

Efficient isolation of cfDNA from 2 ml of human EDTA-treated plasma

Efficient isolation of cfDNA from 2 ml of human EDTA-treated plasma

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.

Back

NucleoSpin Plasma Midi procedure: binding and washing step

NucleoSpin Plasma Midi procedure: binding and washing step

NucleoSpin Plasma Midi procedure: binding and washing step

Back

NucleoSpin Plasma Midi procedure: drying step

NucleoSpin Plasma Midi procedure: drying step

NucleoSpin Plasma Midi procedure: drying step

Back

NucleoSpin Plasma Midi procedure: elution step

NucleoSpin Plasma Midi procedure: elution step

NucleoSpin Plasma Midi procedure: elution step

Back

740302.48: NucleoSpin DNA Plasma Midi Core Kit

740302.48: NucleoSpin DNA Plasma Midi Core Kit
740681 NucleoVac™ 96 Vacuum Manifold Each USD $665.00

NucleoVac 96 Vacuum Manifold vacuum manifold for use of NucleoSpin 8/96 kits under vacuum - manifold base, manifold lid, waste containers, spacer set

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Image Data

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NucleoSpin Tissue purification in less than 1 hour

NucleoSpin Tissue purification in less than 1 hour

NucleoSpin Tissue purification in less than 1 hour. Mouse or rat tail sections, whole tissue, or cultured cells are incubated with lysis buffer and Proteinase K—no further extraction is necessary. After spinning down the lysis mixture, the clear lysates are transferred to the columns and centrifuged to bind the DNA to the column. This is followed by removal of the cellular contaminants with the provided wash buffers, and elution of the purified genomic DNA. No alcohol precipitation is required. The resulting DNA preparation is suitable for PCR, Southern blotting, or any enzymatic reaction. Panel A. NucleoSpin 8 Tissue Kit. Panel B. NucleoSpin 96 Tissue Kit.

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Typical yields of pure genomic DNA from NucleoSpin Tissue range from 15–25 µg

Typical yields of pure genomic DNA from NucleoSpin Tissue range from 15–25 µg

Typical yields of pure genomic DNA from NucleoSpin Tissue range from 15–25 µg. Up to 35 µg of pure genomic DNA can be prepared from two 0.5-cm mouse tail tip sections.

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740681: NucleoVac 96 Vacuum Manifold

740681: NucleoVac 96 Vacuum Manifold

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