QIAamp DSP DNA Blood Mini Kit

For purification of genomic DNA from human whole blood for in vitro diagnostic use

Features

  • Universal DNA purification system compatible with other IVD products
  • Flexible sample collection
  • Compatible with different anticoagulants and tubes
  • Choice of centrifuge protocol or vacuum protocol
QIAamp DSP DNA Blood Mini Kit

Cat. No. / ID: 61104

For 50 preps: QIAamp Mini Spin Columns, Buffers, Reagents, Tubes, VacConnectors
€212.00
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The QIAamp DSP DNA Blood Mini Kit is intended for in vitro diagnostic use.

Product Details

The QIAamp DSP DNA Blood Mini Kit provides silica-based DNA purification. The QIAamp DSP DNA Blood Mini Kit is designed for labs that process blood for in vitro diagnostic use. The procedure can be fully automated on the QIAcube.

Performance

Genomic DNA purified using the QIAamp DSP DNA Blood Mini Kit is ready to use in downstream applications like those based on enzymatic amplification or other modification, such as PCR.

Whole blood samples can be collected using many types of anticoagulant-containing tubes (EDTA or citrate), including BD Vacutainer, Monovette, and Vacuette tubes (see table). Blood samples can be frozen and thawed at least 3 times and still be a reliable source for DNA purification (see figure " DNA from frozen and thawed blood").

Blood collection using a range of different tubes
Primary tube Manufacturer Cat. no. Nominal volumeAverage yield from 200 µl
BD Vacutainer 9NC BD 366007    9 ml 6.4 µg
BD Vacutainer K3E BD 368457  10 ml 6.6 µg
BD Vacutainer K2E BD 367864    6 ml 6.4 µg
S-Monovette EDTA Sarstedt 02.1066.001    9 ml 6.5 µg
S-Monovette CPDA1 Sarstedt 01.1610.001 8.5 ml 6.3 µg
Vacuette K3E Greiner Bio-One 455036    9 ml 6.5 µg
Vacuette 9NC Greiner Bio-One 454382    2 ml 6.3 µg
For each type of primary tube, blood was collected from 11 donors. For each donor, DNA was purified from 200 μl samples in triplicate using the QIAamp DSP DNA Blood Mini Kit and eluted in 200 µl elution buffer. The table shows average DNA yields from 33 purification procedures.

See figures

Principle

The QIAamp DSP DNA Blood Mini Kit uses well-established QIAamp technology for purifying genomic DNA. The QIAamp silica-based membrane specifically binds DNA in the lysed sample, while the rest of the lysate is rapidly removed by either centrifugation or vacuum. The bound DNA is efficiently washed to remove contaminants and then eluted in volumes of 50–200 µl.

Procedure

Genomic DNA is purified from blood using one of 2 alternative procedures, using a centrifuge or using a vacuum manifold and centrifuge (see flowchart " Procedure"). The universal DNA purification system allows compatibility with other in vitro diagnostic products (see " Workflow").

The EDTA- or citrate-containing blood sample (200 µl) is lysed in the presence of QIAGEN Protease and lysis buffer at 56°C for 10 minutes. Ethanol is then added to the lysate to optimize binding of DNA to the QIAamp membrane. The lysate is applied to a QIAamp Mini spin column, which is then centrifuged or subjected to vacuum pressure. DNA binds to the QIAamp membrane in the spin column, and the rest of the lysate passes through. Bound DNA is efficiently washed by 2 different wash buffers, which are also drawn through the QIAamp membrane by centrifugation or vacuum pressure. The QIAamp membrane is then dried by centrifugation. Elution buffer (50–200 µl) is applied to the QIAamp membrane and, after a 1-minute incubation, the spin column is centrifuged to elute pure DNA.

Purification of DNA using the QIAamp DSP DNA Blood Mini Kit can be fully automated on the QIAcube. If automating the QIAamp DSP DNA Blood Mini Kit on the QIAcube instrument, the instrument may process fewer than 50 samples due to dead volumes, evaporation, and additional reagent consumption by automated pipetting. QIAGEN only guarantees 50 sample preps with manual use of the QIAamp DSP DNA Blood Mini Kit.

See figures

Applications

The QIAamp DSP DNA Blood Mini Kit provides proven QIAamp technology for purification of DNA from fresh or frozen whole blood, as well as blood that has been treated with citrate or EDTA.

Supporting data and figures

Specifications

FeaturesSpecifications
ApplicationsPCR, qPCR, real-time RT-PCR, microarray
Main sample typeWhole blood
Elution volume50–200 µl
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinGenomic DNA
CE/FDA/IVD compatibleCE/IVD
FormatSpin columns
Sample amount200 µl
ProcessingManual (centrifugation or vacuum)
TechnologySilica technology
Time per run or per prep20 minutes
Yield6.3–6.5 µg

FAQ

Does the QIAamp DSP Virus Kit require the QIAvac 24 Plus, or can I use it with my own laboratory vacuum system?
It is possible to use a general laboratory vacuum system with the QIAamp DSP Virus Kit. However, this will require validation in-house. The CE-marked QIAamp Kits have been developed, field-tested and validated with the QIAvac 24 Plus vacuum system (QIAvac 24 Plus, QIAvac Connecting System and Vacuum Pump). Using this system with the QIAamp DSP Virus Kit will avoid lengthy validation procedures for the enduser.
FAQ ID -789
What vacuum pressure should be applied using the QIAvac 24 Plus for the QIAamp DSP Virus Kit and Blood Mini Kit Protocols?

When processing samples with the QIAamp DSP DNA Blood Mini Kit and the QIAamp DSP Virus Kit, the vacuum pressure should be below -800 mbar. The QIAvac 24 Plus and the QIAvac Connecting System should be tested according to Appendix B of the QIAvac 24 Plus Handbook before performing a nucleic acid purification procedure.

If you would like to use an equivalent general laboratory vacuum system, make sure that the minimum vacuum pressure is reached. Performance of the QIAamp DSP DNA Blood Mini or Virus Kits in combination with a house-vacuum system has to be validated by the researcher.

FAQ ID -1033
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
What is the difference between QIAGEN Protease and QIAGEN Proteinase K provided in various QIAamp Kits?

QIAGEN Proteinase K is a subtilisin-type protease, which cleaves at the carboxyl side of hydrophobic, aliphatic and aromatic amino acids. It is particularly suitable for short digestion times. It possesses a high specific activity over a wide range of temperatures and pH values with substantially increased activity at higher temperature. Soluble calcium is not essential for enzymatic activity. This means that EDTA, which is used to inhibit Mg2+-dependent enzymes such as nucleases, will not inhibit Proteinase K activity.

QIAGEN Protease is a broad-specificity Serine protease with high activity, cleaving preferentially at neutral and acidic residues. It is an economical alternative to Proteinase K for isolation of native DNA and RNA from a variety of samples.

For more information on activity of QIAGEN Protease and Proteinase K in various buffers please click here.

FAQ ID -761
What can be used as an alternative to the A260 measurement for quantification of small amounts of RNA and DNA?

Small amounts of RNA and DNA may be difficult to measure spectrophotometrically. Fluorometric measurements, or quantitative RT-PCR and PCR are more sensitive and accurate methods to quantify low amounts of RNA or DNA.

Fluorometric measurements are carried out using nucleic acid binding dyes, such as RiboGreen® RNA Quantitation Reagent for RNA, and PicoGreen® DNA Quantitation Reagent for DNA (Molecular Probes, Inc.).

FAQ ID -728
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