PAXgene Blood DNA Kit

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

Features

  • Blood collection and purification in one system
  • Easy handling
  • Enhanced workflow efficiency
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PAXgene Blood DNA Kit (25)

Cat. No. / ID: 761133

Processing tubes and buffers for 25 preparations
$377.00
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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

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 " High quality and high molecular weight of genomic DNA"). 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 " Efficient multiplex PCR of 3 mitochondrial genes").
See figures

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

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

Specifications

FeaturesSpecifications
ApplicationsPCR, qPCR, real-time PCR, blotting, genotyping
ProcessingManual (centrifugation)
Main sample typeWhole blood
Time per run60 min/8 samples
Elution volume1 ml
Yield150–500 µg
FormatProcessing tube (salting out method)
Sample amount8.5 ml

Resources

Brochures & Guides (3)
For collection, storage, and transport of whole blood for genomic DNA isolation

 

Second edition — innovative tools
Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Kit Handbooks (1)
For isolation of genomic DNA from 8.5 ml human whole blood

FAQ

Where can I find additional information for PreAnalytiX PAXgene products?
You can find additional information relating to the PreAnalytiX PAXgene products on the PreAnalytiX website .
FAQ ID - 3515
What does the resuspension buffer, Buffer BG4, in the PAXgene Blood DNA kit contain?

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

FAQ ID - 3501
Does the DNA extracted using the PAXgene Blood DNA System contain RNA?

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
How should blood samples drawn into PAXgene Blood DNA Tubes be transported?

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

FAQ ID - 3507
How long can samples in PAXgene Blood DNA Tubes be stored at room temperature?

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
Are there important considerations for plasma generation and urine handling?

It is strongly advised to follow the recommendations for preparing sample material provided in the corresponding Protocol Sheet to ensure reliable results.

Plasma: It is recommended to perform plasma separation immediately after blood collection when using EDTA or citrate as anticoagulant to prevent the release of genomic DNA into the plasma fraction.

Urine: Because circulating cell-free DNA in non-stabilized urine samples is rapidly degraded after sample collection due to high nuclease activity, eluates may contain no DNA or exhibit low DNA concentration. Therefore, it is recommended to stabilize urine samples. Even when using stabilized urine, it is recommended to perform a centrifugation step immediately after stabilization to prevent the release of genomic DNA from cells. Alternatively, non-stabilized urine samples can be processed immediately after collection and centrifugation using ATL-pretreatment and automated DNA extraction as described in the corresponding Protocol Sheet.

FAQ ID - 3699
3497 - What is the expected purity and size of DNA extracted using the PAXgene Blood DNA kit?

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
Can PAXgene Blood DNA Tubes be stored at higher than room temperature (18–25°C)?

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
What is the expected purity of DNA extracted from blood samples collected in PAXgene Blood DNA Tubes IVD?

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

FAQ ID - 3511
Can mitochondrial DNA be isolated with the PAXgene Blood DNA System?

Yes, mitochondrial and genomic DNA are both isolated.

FAQ ID - 3498
Can smaller volumes of blood be processed from the PAXgene Blood DNA Tube?

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
Which downstream applications have been tested with DNA purified from samples collected in PAXgene Blood DNA Tubes IVD?

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
How should DNA purified using the PAXgene Blood DNA System be stored?

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
Is it possible to interrupt the PAXgene Blood DNA kit protocol?

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
What downstream applications have been tested with DNA purified using the PAXgene Blood DNA System?

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
Can I add bleach to the sample preparation waste?

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
Can I use the PAXgene Blood DNA Tube to collect blood from animals?

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

FAQ ID - 3504
How do I safely inactivate biohazardous flow-through material?

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