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Cat. No. / ID: 51104
✓ 24/7 automatic processing of online orders
✓ Knowledgeable and professional Product & Technical Support
✓ Fast and reliable (re)-ordering
QIAamp DNA Blood Kits provide silica-membrane-based DNA purification from whole blood, plasma, serum and other body fluids. The kits are designed for a range of sample sizes from 200 μl up to 10 ml fresh or frozen human whole blood. QIAamp spin columns can be easily processed in a centrifuge or on vacuum manifolds. A convenient 96-well format using centrifugation enables purification of DNA for labs that need high-throughput DNA purification from blood, buffy coat, plasma, serum, bone marrow, lymphocytes and body fluids. A dedicated kit is also available for automated purification of 1–12 samples on the QIAcube Connect.
QIAamp DNA Blood Kits yield DNA sized from 200 bp to 50 kb, depending on the age and storage of samples (see figure "Apoptotic banding in stored blood"). The purified DNA is suitable for long-range PCR amplification (see figure " Long-range PCR") and restriction fragment length polymorphism (RFLP) analysis used, for example, for paternity testing (see figure " Paternity testing by RFLP analysis").
The QIAamp DNA Blood Midi Kit yields up to 94.5% recovery of DNA; the QIAamp DNA Blood Maxi Kit yields up to 95.8% recovery of DNA, depending on the starting cell densities (see table “DNA yields from human whole blood with different cell densities”).
|Leukocytes per ml||DNA yield (µg)||DNA recovery (%)|
|QIAamp Midi||QIAamp Maxi||QIAamp Midi||QIAamp Maxi|
|2.5 x 105||3.0||15.8||92.3||95.8|
|1.0 x 106||11.0||60.4||91.7||91.5|
|5.0 x 106||62.4||312.0||94.5||94.5|
|1.0 x 107||116.3||624.6||88.1||94.6|
Genomic DNA was purified from 2 ml (QIAamp Midi) or 10 ml (QIAamp Maxi) human whole blood and eluted in 300 µl (Midi) or 1 ml (Maxi) elution buffer. The first eluate was loaded onto the column a second time and centrifuged again (i.e., re-eluted). Percentage DNA recovery was calculated by assuming that one leukocyte contains 6.6 pg DNA.
The QIAamp 96 DNA Blood Kit processes up to 200 µl sample size, with a preparation time of 192 samples in two to three hours, yielding highly pure DNA in less than one minute per preparation. Even higher throughput can be achieved by staggering the procedure. QIAamp 96 plates provide well-to-well uniformity in DNA recovery and purity (see figures " Sample reproducibility" and " Reproducibility of yield and purity"). The typical yield is 6 µg per 200 µl healthy whole blood, with an elution volume of 50–200 µl.
The dedicated QIAamp DNA Blood Mini QIAcube Kit enables automated DNA isolation from blood and DNA isolation from body fluids on the QIAcube Connect. The kit includes rotor adapters that are preloaded with QIAamp spin columns and elution tubes, delivering greater convenience and time savings (see figure " Significant time savings"). Furthermore, ease of use is increased and user errors minimized. Waste is reduced, because the content of the dedicated kit is tailored for purification on the QIAcube Connect and the superfluous tubes that are required for the manual procedure are not included.
If using the automatable QIAamp DNA Blood Mini Kit on the QIAcube Connect, the QIAamp DNA Blood Mini Accessory Set A and QIAamp DNA Blood Mini Accessory Set B provide the convenience of extra buffers and reagents for automated sample prep.
No phenol–chloroform extraction is required. DNA binds specifically to the QIAamp silica-gel membrane while contaminants pass through. PCR inhibitors, such as divalent cations and proteins, are completely removed in two efficient wash steps, leaving pure nucleic acid to be eluted in either water or a buffer provided with the kit. QIAamp DNA Blood technology yields genomic, mitochondrial or viral DNA from blood and related body fluids ready to use in PCR and blotting procedures. QIAamp sample preparation technology is fully licensed.
QIAamp DNA Blood Kits simplify DNA purification from blood and DNA purification from body fluids with fast spin-column, vacuum, centrifugation or automated procedures (see figure “ QIAamp Spin Column procedure”). Fresh and frozen whole blood with common anticoagulants, such as citrate, EDTA and heparin, may be processed. If processing QIAamp Midi or Maxi spin columns on vacuum manifolds additional Buffer AW1 [cat. no. 19081] and Buffer AW2 [cat. no. 19072] are required for use with the vacuum manifolds.
Vacuum processing with the QIAamp DNA Blood Mini Kit
With the QIAamp DNA Blood Mini Kit, blood can be processed by vacuum instead of centrifugation, for greater speed and convenience in DNA purification. QIAamp mini spin columns are accommodated on the QIAvac 24 manifold using VacValves and VacConnectors. VacValves should be used if sample flow rates differ significantly, to ensure consistent vacuum. Disposable VacConnectors are used to avoid any cross-contamination. Use of VacConnectors also allows these QIAamp spin procedures to be performed on QIAvac 6S with QIAvac Luer Adapters.
Automated processing on the QIAcube Connect
The award-winning QIAcube Connect uses advanced technology to process QIAGEN spin columns, enabling seamless integration of automated, low-throughput sample prep into laboratory workflows. All steps in the purification procedure are fully automated – up to 12 samples can be processed per run. The QIAcube Connect used together with the dedicated QIAamp DNA Mini QIAcube Kit provides a winning combination for fast, easy and convenient DNA purification.
High-throughput processing in 96-well format
The QIAamp 96 spin procedure requires the QIAGEN 96-Well-Plate Centrifugation System’s deep rotor buckets to accommodate QIAamp 96 plates stacked on 96-well blocks. Fresh or frozen whole blood treated with common anticoagulants such as, EDTA, citrate and heparin, may be used. Dried whole blood may be processed with additional equipment and the use of a special protocol available from QIAGEN Technical Services or your local distributor.
QIAamp DNA Blood Kits provide proven QIAamp technology for the purification of DNA from a variety of materials. Sample sources include:
|Features||QIAamp DNA Blood Mini Kit||QIAamp DNA Blood Midi Kit||QIAamp DNA Blood Maxi Kit||QIAamp DNA Blood Maxi Kit|
|Applications||PCR, long-range PCR, Southern blotting||PCR, Southern blotting||PCR, blotting||Southern blotting, genome mapping|
|Elution volume||50–200 µl||100–400 µl||500–2000 µl||50–200 µl|
|Format||Spin column||Spin column||Spin column||96-well plate|
|Main sample type||Whole blood, body fluids||Whole blood, body fluids||Whole blood, body fluids||Whole blood, body fluids|
|Processing||Manual (centrifugation or vacuum)||Manual (centrifugation or vacuum)||Manual (centrifugation or vacuum)||Manual (centrifugation or vacuum)|
|Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein||Genomic DNA, mitochondrial DNA, viral DNA||Genomic DNA, mitochondrial DNA, viral DNA||Genomic DNA, mitochondrial DNA, viral DNA||Genomic DNA, mitochondrial DNA, viral DNA|
|Sample amount||1–200 µl||0.3–2 ml||3–10 ml||<200 µl|
|Technology||Silica technology||Silica technology||Silica technology||Silica technology|
|Time per run or per prep||20–40 minutes||55 minutes||55 minutes||2–3 hours (192 samples)|
|Yield||4–12 µg||20–60 µg||300–600 µg||6 µg|
Genomic DNA from 8 blood samples stored at 4°C for 1 week. DNA was purified using the QIAamp DNA Blood Mini Kit. When blood is stored at 4°C, the DNA is rapidly degraded due to apoptosis; the resulting apoptotic banding pattern can clearly be seen in these samples. M1: lambda-HindIII; M2:100 bp ladder.
One milliliter of healthy human blood consists of cell types in approximately the following numbers:
Yes. Please follow the Blood and Body Fluid Spin Protocol in the QIAamp DNA Mini and QIAamp DNA Blood Mini Kit Handbook. No more than 5 x 106 cells should be used. When using the Midi or Maxi format, please follow the protocol for Isolation of DNA using the QIAamp DNA Blood Midi or Maxi Kit in the QIAamp Blood Midi and QIAamp Blood Maxi Handbook. The maximum number of cells to use is 2 x 107 and 1 x 108, respectively.
Yes, we have the following protocols:
For DNA isolation using the QIAamp DNA Mini, or the DNeasy Blood & Tissue Kit, we recommend using carrier DNA when expected yields are below 10 ng. If possible, carrier DNAs such as poly-dA, poly-dT or poly-dA:dT should be used. Other carrier DNAs such as herring sperm DNA may interfere with subsequent PCR by binding primers nonspecifically.
Please note that poly-dA may interfere with oligo-dT primers, and, in this case, a different carrier DNA should be used. The concentration of carrier DNA should be at least 10 µg/ml. Optimal amounts need to be determined empirically for each application. The size distribution of carrier DNAs is typically in the range of 100 bp to 10 kb.
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 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.
Alcohol precipitation is commonly used for concentrating, desalting, and recovering nucleic acids. Since less alcohol is required for isopropanol precipitation, this is the preferred method for precipitation of DNA from large volumes. In addition, isopropanol precipitation can be performed at room temperature, which minimizes co-precipitation of salt that interferes with downstream applications.
Tip: Use a buffer with a pH of 7.5–8.0, as DNA does not dissolve easily in acidic buffers. Often distilled water can have an acidic pH. The addition of EDTA protects the DNA from DNase digestion.
Tip: High-molecular-weight DNA, such as genomic DNA, should be redissolved very gently to avoid shearing. If the DNA pellet does not dissolve easily, heat at 55°C for 1–2 h with gentle shaking.
Yes, we please follow the Supplementary Protocol 'Isolation of genomic DNA from dried blood spots using the QIAamp 96 DNA Blood Kit' (QA22). Please contact Technical Service for this protocol.
Yes, please follow the User-Developed Protocol 'Isolation of bacterial DNA from soil using the QIAamp DNA Stool Mini Kit and QIAamp DNA Blood Midi Kit' (QA28).
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.
Yes. We have shown that DNA purified with the QIAamp DNA Blood Mini Kit is stable for at least 10 years at either 2–8ºC or –20ºC. However, our data indicates that DNA stability is dependent upon elution buffer used for storage. For detailed information on this stability study see the QIAGEN News Article for the first part of the study and then the continuing study data at the 10 year mark.
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.).
See: http://www.qiagen.com/Products/Catalog/Automated-Solutions/Sample-Prep/QIAcube#orderinginformation for a virtual demonstration on how to use the QIAcube.
The composition of Buffer AE is:
According to an Interview with Professor Dennis Lo published in QIAGEN News Molecular Diagnostics, Issue No. 5, 2002, healthy individuals have about 500-1000 genome equivalents (DNA) per ml serum/plasma.
For free-circulating DNA in plasma, the concentration can range from 1–100 ng/ml in healthy individuals.
Tissue is more difficult to lyse than cells in a blood sample. Therefore an additional incubation step in buffer ATL containing Proteinase K is included in the Tissue Protocol to achieve complete sample lysis. Blood cells will directly lyse by incubation in Buffer AL containing QIAGEN Protease. Buffer AL is required in both protocols to ensure appropriate conditions for DNA binding to the silica membrane. For further details on the protocols, please refer to the QIAamp DNA Mini and QIAamp DNA Blood Mini Kit Handbook.
QIAGEN Protease is inactivated by incubation at 70°C for 15 minutes.
To our knowledge, Proteinase K cannot be completely heat-inactivated. Even when incubating at 95°C for 10 minutes, some enzymatic activity remains. This will not negatively affect the QIAamp Procedure, since the enzyme will be efficiently removed by the wash steps in the protocols.
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.
EDTA chelates divalent cations which are required for nuclease activity. While the genomic DNA (gDNA) extracted using QIAGEN products, should not have any nuclease activity, it is possible to introduce nucleases during repeated long-term access of the DNA. EDTA helps to prevent any nuclease activity introduced after the genomic DNA extraction procedures.
However, if the gDNA is stored frozen at -20oC or -80oC, nuclease activity is much reduced. It may be possible to leave EDTA out of the storage buffer without negative consequences when samples are kept under these conditions, and when repeated freeze-thaw cycles are avoided.
We do recommend however that gDNA be stored in a neutral to a slightly basic buffered solution (e.g. 10 mM Tris-Cl pH 8.5 to 9.0) to prevent DNA degradation by acid hydrolysis. Note that deionized water mostly has an acidic pH.