PAXgene Blood DNA Kit

For purification of up to 500 µg genomic DNA from whole blood

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PAXgene Blood DNA Kit (25)

Cat. No. / ID: 761133

Processing tubes and buffers for 25 preparations

For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information 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

Blood collection and purification in one system
Easy handling
Enhanced workflow efficiency

Product Details

The PAXgene Blood DNA system consists of PAXgene Blood DNA Tubes for blood collection and stabilization, and the PAXgene Blood DNA Kit for DNA purification in a single-tube procedure.

Performance

DNA isolated using the PAXgene Blood DNA System has an A₂₆₀/A₂₈₀ ratio of 1.7–1.9. Fragments are 20–200 kb in size, with an average length of 50–150 kb (see figure ""). Average DNA yields are 150–500 µg, depending on the number of nucleated cells present in the blood sample. Purified DNA can be used in PCR and quantitative, real-time PCR (see figure "").

See figures

High quality and high molecular weight of genomic DNA.

Efficient multiplex PCR of 3 mitochondrial genes.

Principle

The PAXgene Blood DNA System is an integrated and standardized system for collection and stabilization of whole blood specimens and isolation of their genomic DNA. The system requires the combined use of PAXgene Blood DNA Tubes for blood collection and stabilization, followed by DNA isolation using the PAXgene Blood DNA Kit.

Blood is collected into PAXgene Blood DNA Tubes, which contain a proprietary blend of reagents that both prevents blood coagulation and stabilizes white blood cells. Blood samples can be stored in the tubes for up to 14 days at room temperature. DNA is isolated from the tubes using appropriate buffers supplied in the PAXgene Blood DNA Kit. Highly pure genomic DNA is obtained, suitable for use in a wide range of downstream applications.

Procedure

The PAXgene Blood DNA procedure is shown in the flowchart " PAXgene Blood DNA procedure". Blood samples (8.5 ml) are collected in PAXgene Blood DNA Tubes, where they may be stored or transported. For DNA isolation, the blood is transferred to processing tubes (supplied already filled with cell lysis buffer), and the solution is mixed to lyse red and white blood cells. Cell nuclei and mitochondria are pelleted by centrifugation, washed, and resuspended in digestion buffer. Protein contaminants are removed by incubation with a protease. DNA is precipitated in isopropanol, washed in 70% ethanol, dried, and resuspended in resuspension buffer.

See figures

PAXgene Blood DNA procedure.

Applications

The DNA can be used in a wide range of downstream applications, including:

PCR and quantitative, real-time PCR
Southern blotting
SNP genotyping
Pharmacogenomic studies
SNP discovery and Validation

Supporting data and figures

PAXgene Blood DNA procedure.

Blood samples (8.5 ml) are collected in PAXgene Blood DNA Tubes, where they may be stored or transported. For DNA isolation, the blood is transferred to processing tubes (supplied already filled with cell lysis buffer), and the solution is mixed to lyse red and white blood cells. Cell nuclei and mitochondria are pelleted by centrifugation, washed, and resuspended in digestion buffer. Protein contaminants are removed by incubation with a protease. DNA is precipitated in isopropanol, washed in 70% ethanol, dried, and resuspended in resuspension buffer.

Specifications

Features	Specifications
Main sample type	Whole blood
Time per run	60 min/8 samples
Elution volume	1 ml
Yield	150–500 µg
Format	Processing tube (salting out method)
Sample amount	8.5 ml
Applications	(RT-)PCR, (RT-)qPCR, NGS, genotyping, microarray analysis
Processing	Manual (centrifugation)

Resources

Second edition — innovative tools

For collection, storage, and transport of whole blood for genomic DNA isolation

Groelz et al., ASHG 2004

For isolation of genomic DNA from 8.5 ml human whole blood

FAQ

You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .

FAQ ID - 3515

Buffer BG4 consists of 10 mM Tris·Cl, pH 8.5.

FAQ ID - 3501

No. DNA purified with the PAXgene Blood DNA System is free of detectable RNA contamination. In contrast to commercial salting out methods, additional RNase digestion steps are not necessary.

FAQ ID - 3499

See subsequent FAQs for information about storage interval and temperature. Each laboratory should validate shipping procedures.

FAQ ID - 3507

Samples in PAXgene Blood DNA Tubes can be stored at room temperature (18–25°C) for up to 14 days after blood collection.

FAQ ID - 3508

The typical A260/A280 ratio is from 1.7 to 1.9. The DNA molecules are up to 200 kb in size, with fragments of 50–150 kb predominating.

FAQ ID - 3497

Yes, samples in PAXgene Blood DNA Tubes can be stored at 30°C for up to 3 days after blood collection. However, it is not recommended to store blood samples at temperatures higher than 30°C and storage at high temperatures should not exceed 3 days.

FAQ ID - 3509

The typical A₂₆₀/A₂₈₀ ratio is from 1.7 to 1.9.

FAQ ID - 3511

Yield of genomic DNA varies depending on the white blood cell count of the patient blood sample, the extraction method, and the elution volume. However, DNA yield typically ranges between 16–135 µg/ml whole blood. Specifically, DNA yields from magnetic bead-based extraction on the QIAsymphony ranges between 16.9–51.7 µg/ml whole blood, from silica membrane-based extraction on the QIAcube 17.1–45.8 µg/ml whole blood, and from salting-out precipitation 4.9–53.7 µg/ml whole blood.

FAQ ID - 3510

Yes, mitochondrial and genomic DNA are both isolated.

FAQ ID - 3498

No. The volume of Buffer BG1 (lysis buffer) prefilled in the processing tubes, and all the other buffer volumes are optimized for use with blood from one PAXgene Blood DNA Tube.

FAQ ID - 3500

To date, DNA purified from blood samples in PAXgene Blood DNA Tubes has been successfully tested for use in PCR, including multiplex PCR.

FAQ ID - 3512

DNA purified using the PAXgene Blood DNA System should be dissolved in Buffer BG4 and stored at 2–8°C. For long-term storage, DNA can be stored at –20°C but multiple freezing and thawing cycles should be avoided.

FAQ ID - 3502

Yes. After dissolving the nuclei pellet in digestion buffer, Buffer BG3 (step 7), samples can be stored up to 7 days at 2–8°C or for one day at room temperature (18–25°C). The protocol can also be interrupted after adding isopropanol (step 10).

FAQ ID - 3503

So far, we have shown that DNA purified using the PAXgene Blood DNA System can be used in PCR, including long range and multiplex PCR, restriction digestion and Southern analysis, as well as in various SNP detection procedures including,TaqMan®, LightCycler and Masscode technologies.

See trademarks.

FAQ ID - 3506

Do not add bleach or acidic solutions directly to the sample preparation waste. CAUTION: The sample preparation waste contains guanidinium hydrochloride from Buffer BG3, which can form highly reactive compounds when combined with bleach. For more information please consult the appropriate material safety data sheets (MSDS).

FAQ ID - 3505

The system is designed for use only with human blood samples.

FAQ ID - 3504

Always dispose of potentially biohazardous solutions according to your institution’s waste-disposal guidelines. Although the lysis and binding buffers in QIAamp, DNeasy, and RNeasy kits contain chaotropic agents that can inactivate some biohazardous material, local regulations dictate the proper way to dispose of biohazards. DO NOT add bleach or acidic solutions directly to the sample-preparation waste. Guanidine hydrochloride in the sample-preparation waste can form highly reactive compounds when combined with bleach.
Please access our Material Safety Data Sheets (MSDS) online for detailed information on the reagents for each respective kit.

FAQ ID -12