PAXgene Blood DNA Tube
Sample Collection and Phlebotomy
1. Do I have to use a blood collection set with the PAXgene Blood DNA Tube?
No, a blood collection set does not have to be used with the PAXgene Blood DNA Tube. Further information on phlebotomy technique with the PAXgene Blood DNA Tube can be found in the Handbook.
2. Are there any special considerations for phlebotomy when using the PAXgene Blood DNA Tube (blood collection set, discard tube, positioning of the tube, issues with short draws, etc.).
No, blood collection is performed under a standard phlebotomy protocol into the evacuated tube using a needle or blood collection set and a standardized needle holder that can accommodate a 16 mm diameter tube. For more information on phlebotomy technique, please refer to the PAXgene Blood DNA Tube Handbook.
3. What is the reagent in the tube?
The PAXgene Blood DNA Tube contains a proprietary EDTA formulation. Contents of the tube may cause irritation to eyes, respiratory system and skin. Please refer to the PAXgene Blood DNA Tube Handbook or for additional information, visit the PreAnalytiX website or contact QIAGEN Technical Service to obtain the SDS sheet.
4. Are the PAXgene Blood DNA Tubes sterile?
The inside of the PAXgene Blood DNA Tube is sterilized during production.
5. What is the shelf life of the PAXgene Blood DNA Tube prior to blood collection?
The expiration date is printed on the tube label and on the shelf carton. Store the PAXgene Blood DNA Tubes at room temperature (18–25°C).
1. How should blood samples drawn into PAXgene Blood DNA Tubes be shipped?
See subsequent FAQs for specific storage temperature and time information. Individual laboratories should validate shipping procedures.
Sample Storage and Archiving
1. How should blood samples drawn into the PAXgene Blood DNA Tubes be stored?
Blood samples can be stored at 15–25°C for up to 14 days or at 2–8°C for up to 28 days. For longer-term storage, freezing the samples at –70ºC or –80°C is recommended.
2. Can the PAXgene Blood DNA Tube be frozen? If yes, at what temperatures and for how long?
The PAXgene Blood DNA Tubes can be stored at –20ºC for up to 3 months. If tubes are to be kept at temperatures below –20ºC, freeze them first at –20ºC for 24 hours, then transfer them to –70ºC or –80ºC.
To freeze PAXgene Blood DNA Tubes, stand them upright in a wire rack or horizontally in a plastic bag. Do not freeze tubes upright in a styrofoam tray as this may cause the tubes to crack.
3. How long can the PAXgene Blood DNA Tube be kept at room temperature? What is room temperature?
The PAXgene Blood DNA Tubes can be kept at room temperature for up to 14 days after blood collection. Room temperature is designated as 18–25°C.
4. Can the PAXgene Blood DNA Tubes be stored at 4°C?
Yes, the PAXgene Blood DNA Tubes can be stored at 4°C for up to 28 days after blood collection.
5. Can the PAXgene Blood DNA Tubes be stored at higher temperatures?
Yes, the PAXgene Blood DNA Tubes can be stored at 30°C for up to 7 days after blood collection. However it is not recommended to store blood sample at temperatures higher than 30°C.
DNA Isolation and Processing
1. How should I thaw frozen PAXgene Blood DNA Tubes?
Thaw the PAXgene Blood DNA Tubes in a wire rack at ambient temperature (18–25ºC) for approximately two hours or at up to 37°C in a water bath for approximately 15 minutes. Carefully invert the thawed PAXgene Blood DNA Tubes 10 times.
2. What is the expected yield from 8.5 ml of blood?
Yield of genomic DNA will vary depending on the cell count from the donor blood sample but average range is from 150 µg to 500 µg.
3. What is the expected purity and size of DNA?
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.
4. Can mitochondrial DNA be isolated with the PAXgene Blood DNA System?
Yes, mitochondrial and genomic DNA are both isolated.
5. Does the PAXgene Blood DNA System contain RNA contamination?
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.
6. 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.
7. What does the resuspension buffer, Buffer BG4, contain?
Buffer BG4 consists of 10 mM Tris·Cl, pH 8.5.
8. 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.
9. Is it possible to interrupt the 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).
10. Can I use the PAXgene Blood DNA Tube to collect blood from animals?
The system is designed for use only with human blood samples.
11. 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 safety data sheets (SDS).
1. 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.