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PAXgene Blood DNA Kit

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

  • Efficient purification of high-quality, high-molecular weight DNA
  • Optimized DNA purification from specimens collected in PAXgene Blood DNA Tubes
  • Rapid procedure in a single processing tube
  • Standardized and easy workflow
     

Feature

Specification

Format  

Processing tube

Technology 

Lysis, digestion, and precipitation

Sample input volume 

Content of one PAXgene Blood DNA Tube

Elution volume

1 mL

Sample type

8.5 ml whole blood, drawn into a PAXgene Blood DNA Tube

Time per run

60 min/8 samples

Typical DNA yield 

150–500 µg

DNA purity 

1.7–1.9 (A260/A280)

DNA size  

50–200 kb

Processing  

Manual: centrifugation


Intended Use

The PAXgene Blood DNA Kit is intended for the purification of DNA from whole blood samples. It is intended to be used as part of the PAXgene Blood DNA System, which additionally includes the PAXgene Blood DNA Tube for the collection and storage of whole blood.

For research use only. Not for use in diagnostic procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease. The performance characteristics of this product have not been fully established.

Order information

Product

Catalog No.

Price

PAXgene Blood DNA Kit (25)

Processing tubes and buffers for 25 preparations. To be used in conjunction with PAXgene Blood DNA Tubes.

761133

Details

Performance

DNA from whole blood samples collected in PAXgene Blood DNA Tubes purified using the PAXgene Blood DNA Kit has an A260/A280 ratio of 1.7–1.9. Fragments range between 20–200 kb in size, with an average length of 50–150 kb (Figure 1). Average DNA yields are 150–500 µg, depending on the number of nucleated cells present in the blood sample.

Principle

Applications such as pharmacogenomic studies, gene expression studies, and SNP genotyping yield clinically important data and can require large amounts of purified DNA from individual subjects. Use of standardized methods for sample collection and nucleic acid isolation to produce high-quality DNA for such studies is becoming increasingly important for multi-center clinical trials and basic research. The purification process for the PAXgene Blood DNA Kit is performed in a single processing tube to minimize the risk of sample mix-up and cross-contamination. Together with the PAXgene Blood DNA Tube, which standardizes the collection and storage of whole blood at various temperatures, these two components of the PAXgene Blood DNA System comprise a simplified, integrated and standardized preanalytic workflow for blood samples used in DNA studies.

Procedure

The procedure to purify DNA using the PAXgene Blood DNA Kit is shown in the flowchart (Figure 2). Briefly, blood samples collected in PAXgene Blood DNA Tubes are 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.

Applications

The resulting high-quality DNA can be used for demanding downstream applications, including:

  • PCR, multiplex, long-range, and quantitative, real-time PCR
  • Southern blotting
  • SNP genotyping
  • Next-generation sequencing
     
Genomic DNA isolated from 8 blood donors using the PAXgene Blood DNA System. [A] Agarose gel analysis; [B] pulsed-field gel electrophoresis for enhanced separation of high-molecular-weight genomic DNA. M: markers.

Figure 1. High quality and high molecular weight of genomic DNA.

Genomic DNA isolated from 8 blood donors using the PAXgene Blood DNA System. [A] ...
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.

Figure 2. PAXgene Blood DNA procedure.

Blood samples (8.5 ml) are collected in PAXgene Blood DNA Tubes, where they may be ...
Multiplex PCR of fragments from the mitochondrial genes tRNAlys/ATPase (0.92 kb), tRNAleu(UUR) (0.63 kb), and ND4 (0.29 kb), using 250 ng DNA from 8 donors as starting material.

Figure 3. Efficient multiplex PCR of 3 mitochondrial genes.

Multiplex PCR of fragments from the mitochondrial genes tRNAlys/ATPase (0.92 kb), ...
Multiplex PCR from gene fragments CD19 (955 bb), CD59 (845 bp), CD40 (756 bp), CD14 (662 bp), AGTR (523 bp), cKit (414 bp), B29 E4 (310 bp) and PRP (222 bp), using 250 ng DNA isolated from 8 blood donors using the PAXgene Blood DNA System as starting material. M: markers.

Figure 4. Efficient multiplex PCR amplification of 8 fragments from single copy genes.

Multiplex PCR from gene fragments CD19 (955 bb), CD59 (845 bp), CD40 (756 bp), ...
Amplification of a 15 kb fragment of the human coagulation factor IX gene, using 250 ng DNA isolated from 8 blood donors using the PAXgene Blood DNA System as starting material. M: markers.

Figure 5. Efficient amplification of a 15 kb gene fragment.

Amplification of a 15 kb fragment of the human coagulation factor IX gene, using ...

Resources

(EN) Important Note: PreAnalytiX GmbH street address has changed from “Feldbachstrasse” to “Garstligweg 8”

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Effect of preanalytical factors on analyte quality as shown with the QIAxcel Connect capillary gel electrophoresis system

1.7 MB

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Other Products Used in Conjunction

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