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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.
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").
Primary tube | Manufacturer | Cat. no. | Nominal volume | Average 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 |
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.
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.
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.
EDTA-treated blood samples were frozen and thawed up to 3 times. DNA was purified at the end of each freeze-thaw cycle. DNA was purified from 200 µl samples using the QIAamp DSP DNA Blood Mini Kit and eluted in 200 µl elution buffer. DNA yields are normalized to the yield from fresh sample (100%). Each bar represents the results from 32 replicates (mean ± standard deviation).
Features | Specifications |
---|---|
Applications | Downstream detection procedures in molecular diagnostics, such as PCR |
Main sample type | Whole blood |
Elution volume | 50–200 µl |
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein | Genomic DNA |
CE/FDA/IVD compatible | CE/IVD |
Format | Spin columns |
Sample amount | 200 µl |
Processing | Manual (centrifugation or vacuum); automated (on QIAcube Connect MDx) |
Technology | Silica technology |