QIAamp MinElute ccfDNA Kits

For isolation of free-circulating DNA from human plasma or serum

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QIAamp MinElute ccfDNA Midi Kit (50)

Cat. No. / ID: 55284

For 50 preps (4 or 5 ml sample input volume each): QIAamp UCP MinElute Columns, QIAGEN Proteinase K, Magnetic Bead Suspension, Buffers, Bead Elution Tubes, and Collection Tubes (1.5 ml and 2 ml)

KitAccessories

QIAamp MinElute ccfDNA Kit

Accessories

Volume

5–10 ml

1–4 ml

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QIAamp MinElute ccfDNA Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

Efficient recovery of fragmented DNA
High yield ensuring detection of low frequency biomarkers
Low elution volume in water (20 µl) for high concentration of nucleic acids
Scalable sample input volume of 1–10 ml
Removal of contaminants and inhibitors

Product Details

QIAamp MinElute ccfDNA Kits greatly simplify concentration and purification of free-circulating DNA from plasma or serum. The procedure can be partially automated on the QIAcube Connect.

Performance

The quantity of free-circulating DNA in plasma, serum and other body fluids is usually low, and isolation is a challenge. QIAamp MinElute ccfDNA Kits provide excellent results with efficient purification of high-quality ccfDNA from serum and plasma samples, even when small amounts are present. Compared with other commercially available kits, QIAamp MinElute ccfDNA Kits recovered the greatest amount of ccfDNA.
Pre-concentration of ccfDNA on magnetic beads allows flexible and scalable handling of volumes from 1 to 10 ml while the use of a MinElute column allows flexible elution volumes between 20 μl and 80 μl, providing the highest possible concentration of nucleic acid species that are present in low concentrations (typically 1–100 ng/ml circulating DNA in human plasma).
The QIAamp MinElute ccfDNA Mini Kit has been optimized for sample volumes from 1 to 4 ml while the QIAamp MinElute ccfDNA Midi Kit has been optimized for sample volumes from 4 to 10 ml. QIAamp MinElute columns can bind fragmented nucleic acids that are as short as 20 bases, but yield depends on the sample volume and the concentration of circulating nucleic acids in the sample.

Principle

Free-circulating DNA, such as tumor-specific extracellular DNA fragments in the blood or fetal DNA in maternal blood, is present in serum or plasma usually as short fragments of <1000 bp. Because ccfDNA may reflect the entire tumor genome, it has gained attention for its potential clinical utility, enabling use of “liquid biopsies” in the form of blood draws.
QIAamp MinElute ccfDNA Kits enable efficient purification of these circulating DNAs from human plasma or serum. The procedure can be partially automated on the QIAcube Connect.

Procedure

The kits use a fast procedure involving pre-concentration of circulating nucleic acids onto magnetic beads and a cleanup of the resulting pre-eluate on QIAamp MinElute columns in a quick and convenient spin procedure. The robust procedure helps to eliminate sample-to-sample cross-contamination and increases user safety when handling potentially infectious samples.
The simple procedure, which is highly suited for simultaneous processing of multiple samples, provides pure nucleic acids in less than 1 hour for 24 samples. Free-circulating DNA is eluted in Ultra-clean Water, ready for use in downstream reactions or storage a –30 to –15°C. Purified nucleic acids are free of proteins, nucleases and other impurities. The procedure can be partially automated on the QIAcube.

Applications

QIAamp MinElute ccfDNA Kits enable efficient purification of circulating DNAs from human plasma or serum.

Supporting data and figures

Faster access to ccfDNA

Comparison of sample processing time (for 4 samples of 4 ml plasma each, using commercially available kits) shows that the QIAamp MinElute ccfDNA Kit provided the fastest processing.

Resources

Digital PCR (dPCR) is a powerful technique that detects and quantifies ultra-rare mutations in a high background of wild-type cfDNA down to 0.1% variant allele frequency. Here, we describe end-to-end manual and automated workflows that enable accurate detection and absolute quantification of ultra-rare PIK3CA variants in cfDNA using the QIAcuity Digital PCR System.

FAQ

The QIAsymphony PAXgene Blood ccfDNA Kit can be sued with 2 protocol lines. The standard protocols (STA) predominantely isolate small ccfDNA fragments. The large fragment protocols (LAF) efficiently co-purify large and small ccfDNA fragments. Both lines include a protocol for 2.4 ml and another for 4.8 ml plasma input.

FAQ ID - 3654

The elution volume of the standard protocols (STA) is fixed at 60 µl. The large fragment protocols (LAF) enable elution in 3 different volumes: 60, 100 and 150 µl.

FAQ ID - 3655

The expiration date is printed on the kit label and encoded in the bar code of the reagent cartridges.

FAQ ID - 3651

For overview and further information on the Globally Harmonized System of Classification and Labelling of Chemicals (GHS) please visit www.qiagen.com/ghs .

FAQ ID - 3428

Whole blood drawn into PAXgene Blood ccfDNA Tubes can be stored for at least 24 hours at 35°C. Higher temperatures will lead to lysis of blood cells and release of genomic DNA into plasma.

FAQ ID - 3640

To date, ccfDNA from blood samples collected and processed using the PAXgene Blood ccfDNA System have been successfully tested in PCR, including multiplex qPCR, and NGS applications.

FAQ ID - 3658

The recommended sample input format is the 14 ml Falcon® polystyrene round bottom tube 17 x 100 mm supplied by Corning (formerly supplied by BD).

FAQ ID - 3653

No. Freezing blood drawn into PAXgene Blood ccfDNA Tubes results in lysis of red and white blood cells and release of genomic DNA.

FAQ ID - 3642

Yes, the inside of PAXgene Blood ccfDNA Tubes is sterilized during production.

FAQ ID - 3636

The expiration date is printed on the tube label and on the shelf carton.

FAQ ID - 3633

When closed properly with the provided strips and stored at room temperature (15–25°C), a reagent cartridge can be used up to 6 weeks after the initial run.

FAQ ID - 3650

Yes. Whole blood in PAXgene Blood ccfDNA Tubes can be stored at 8–15°C for up to 7 days. Storing blood in PAXgene Blood ccfDNA Tubes below room temperature does not extend stability.

FAQ ID - 3641

If cryoprecipitates cannot be resolved completely by vortexing, the sample should be centrifuged to prevent precipitates from blocking the column. To ensure that no cfDNA is co-precipitated, centrifugation can be performed at a lower speed (at 6,800xg for 3 minutes at room temperature, as stated in the Protocol Sheet for the QS DSP Circulating DNA Kit).

FAQ ID - 3646

Plasma generated from whole blood collected into PAXgene Blood ccfDNA Tubes can be stored at 2–8°C for up to 14 days. We recommend freezing plasma at −20°C or −80°C for longer storage periods (long-term storage studies are ongoing).

FAQ ID - 3644

The yield of ccfDNA depends on the target ccfDNA. Nevertheless, yields obtained from PAXgene Blood ccfDNA Tubes are comparable to those obtained from the same volume of plasma separated directly after drawing blood into a spray-dried EDTA tube. In the PAXgene Blood ccfDNA Tube, however, this yield remains stable for up to 7 days when the tube is stored at 15–25°C or for up to 24 hours when stored at 35°C. In contrast, genomic DNA is released from lysed and apoptotic cells in the unstabilized contents of the EDTA tube soon after blood draw.

FAQ ID - 3637

Yes. Further information on phlebotomy technique with the PAXgene Blood ccfDNA Tube can be found in the PAXgene Blood ccfDNA Tube Product Circular under the Resources tab.

FAQ ID - 3626

The number of extractions that can be performed with a kit is independent of sample volume. A kit contains sufficient reagents for 192 extractions.

FAQ ID - 3649

If the instructions for use are followed, there is no possibility of backflow during phlebotomy and thus, no possibility of reagent going into the patient’s arm.

FAQ ID - 3629

A complete 10 ml blood draw ensures optimal results. However, depending on the downstream application and target ccfDNA fragment size, tubes filled with as little as 6.5 ml whole blood can be used without adversely affecting results.

FAQ ID - 3638

Yes, as long as the phlebotomy instructions included with the PAXgene Blood ccfDNA Tubes are properly followed.

FAQ ID - 3630

The PAXgene Blood ccfDNA Tube contains a proprietary formulation that prevents blood coagulation and the release of genomic DNA from blood cells into plasma. Contents of the tube may cause irritation to the eyes, respiratory system and skin. Please refer to the PAXgene Blood ccfDNA Tube Product Circular. For additional information, visit www.preanalytix.com or review the Safety Data Sheet under the Resources tab.

FAQ ID - 3632

The blood draw volume is 10 ml and the stabilization reagent volume is 1.5 ml. Thus, a filled tube holds a volume of 11.5 ml.

FAQ ID - 3631

The QIAsymphony PAXgene Blood ccfDNA Kit is designed to be used with plasma generated from whole blood collected into PAXgene Blood ccfDNA Tubes.

FAQ ID - 3652

Thaw the tubes at room temperature (15–25°C). To minimize the formation of cryoprecipitates, do not thaw at lower temperatures (e.g., 4°C). Alternatively, thaw plasma for 30 minutes at 30°C to completely avoid the formation of cryoprecipitates.

FAQ ID - 3645

Yes. Following a standard phlebotomy protocol, blood is collected into the evacuated tube using a blood collection set and a standardized needle holder that can accommodate a 16 mm diameter tube. Make sure that blood has stopped flowing into the tube before removing the tube from the holder.

For more information on phlebotomy technique, please refer to the PAXgene Blood ccfDNA Tube Product Circular under the Resources tab.

FAQ ID - 3635

No. However, a second centrifugation step helps remove traces of buffy coat that may be accidentally transferred when aspirating plasma after the first centrifugation step.

FAQ ID - 3643

Eluates should be stored at −20°C (long-term storage studies are ongoing).

FAQ ID - 3657

Yes, a customized protocol for elution with 50 µl is available on request.

FAQ ID - 3656

Since the Qproteome Plasma Membrane Protein procedure requires 1.5 ml/2.0 ml reaction tubes, we recommend using the follwing magnets for handling the magnetic beads:

QIAGEN’s Single-Tube Magnet
QIAGEN’s 12-Tube Magnet

Alternatively, comparable tube magnets or a standard clip-board magnet should also work.

FAQ ID -1193

Whole blood drawn into PAXgene Blood ccfDNA Tubes can be stored at 15–25°C for up to 7 days.

FAQ ID - 3639

The Globally Harmonized System (GHS) is an initiative by the United Nations aiming to harmonize chemical hazard communication worldwide. Hazards symbols accompanying products are changing to those of the GHS. Information on the adoption of this new system is available on our GHS Hazard Labeling pages.

FAQ ID - 3430

The normal protocols enable processing 2.4 or 4.8 ml plasma. Other volumes can be accommodated using customized protocols.

FAQ ID - 3648

Store PAXgene Blood ccfDNA Tubes at room temperature (15–25°C) and protect them from direct sunlight.

FAQ ID - 3634

When used in accordance with instructions, the blood collection set eliminates the possibility of patient contact with the reagent in the tube due to backflow of blood.

FAQ ID - 3628