QIAamp DNA Blood Kits – Genomic DNA Extraction

For purification of genomic, mitochondrial or viral DNA from blood and other body fluids

S_1433_RPA_QA0943

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QIAamp DNA Blood Mini Kit (50)

Cat. No. / ID:  51104

For 50 DNA minipreps: 50 QIAamp Mini Spin Columns, QIAGEN Protease, Reagents, Buffers, Collection Tubes (2 ml)
KitAccessories
QIAamp DNA Blood Kit
QIAamp DNA Blood Mini Accessory Set
Column typePlate type
Mini
Mini QIAcube
Midi
Maxi
96 well
Preparations
50
250
QIAamp DNA Blood Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Rapid purification of high-quality, ready-to-use DNA
  • No organic extraction or alcohol precipitation
  • Consistent, high yields
  • Complete removal of contaminants and inhibitors for reliable results
  • Kit formats for low- to high-throughput – options for automation of all kits

Product Details

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.

Performance

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”).

 

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.

See figures

Principle

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.

Procedure

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.

See figures

Applications

QIAamp DNA Blood Kits provide proven QIAamp technology for the purification of DNA from a variety of materials. Sample sources include:

  • Fresh and frozen whole blood or buffy coat
  • Plasma or serum
  • Bone marrow
  • Lymphocytes
  • Platelets
  • Body fluids
  • Cultured cells
  • Swabs and buccal cells

 

Comparison of QIAamp DNA Blood Kits
Features QIAamp DNA Blood Mini Kit QIAamp DNA Blood Midi Kit QIAamp DNA Blood Maxi Kit QIAamp 96 DNA Blood 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

Supporting data and figures

Resources

Kit Handbooks (6)
For large-scale genomic and viral DNA purification from whole blood, plasma, serum, body fluids, lymphocytes
User-Developed Protocols (2)
As starting material, 5 g soil was mixed with different amounts of Bacillus subtilis cells. Sensitivity was 5 x 103 cells/5g soil.
Brochures & Guides (5)
Second edition — innovative tools
Introducing QIAseq
PDF (450KB)
Accelerate your NGS performance through Sample to Insight solutions
Safety Data Sheets (1)
Quick-Start Protocols (1)
Technical Information and Important Notes (2)
Certificates of Analysis (1)
Request a Certificate of Analysis and see the quality control data for your product.

Publications

A multiplex ligase detection reaction-fluorescent microsphere assay for simultaneous detection of single nucleotide polymorphisms associated with Plasmodium falciparum drug resistance.
Carnevale EP; Kouri D; DaRe JT; McNamara DT; Mueller I; Zimmerman PA;
J Clin Microbiol; 2006; 45 (3):752-61 2006 Nov 22 PMID:17121999
The association of sequence variants in DNA repair and cell cycle genes with cancers of the upper aerodigestive tract.
Hall J; Hashibe M; Boffetta P; Gaborieau V; Moullan N; Chabrier A; Zaridze D; Shangina O; Szeszenia-Dabrowska N; Mates D; Janout V; Fabiánová E; Holcatova I; Hung RJ; McKay J; Canzian F; Brennan P;
Carcinogenesis; 2006; 28 (3):665-71 2006 Oct 13 PMID:17040931
Individual-based assessment of population structure and admixture in Austrian, Croatian and German draught horses.
Druml T; Curik I; Baumung R; Aberle K; Distl O; Sölkner J;
Heredity (Edinb); 2006; 98 (2):114-22 2006 Oct 11 PMID:17035951
Quantification of genital human immunodeficiency virus type 1 (HIV-1) DNA in specimens from women with low plasma HIV-1 RNA levels typical of HIV-1 nontransmitters.
Benki S; McClelland RS; Emery S; Baeten JM; Richardson BA; Lavreys L; Mandaliya K; Overbaugh J;
J Clin Microbiol; 2006; 44 (12):4357-62 2006 Oct 18 PMID:17050820

FAQ

Can I use your QIAvac 96 for extraction of the DNA using QIAamp 96 DNA Blood Kit?

We do not recommend using a vacuum manifold for DNA extraction using the QIAamp 96 DNA Blood Kit because:

  1. Viscosity of the lysates varies from sample to sample. Therefore, the wells of the QIAamp 96 plate are at great risk of clogging.
  2. Use of vacuum manifold would result in incomplete removal of ethanol and salts after washing with Buffer AW2. This will affect the purity and yield of samples.
FAQ ID -9158
What is the cellular composition of human blood?

One milliliter of healthy human blood consists of cell types in approximately the following numbers:

  • Leucocytes (function: immune response) 4–7 x 106 cells
  • Thrombocytes (function: wound closing) 3–4 x 108 cells 
  • Erythrocytes (function O2 and CO2 transport) 5 x 109 cells
FAQ ID -2951
Do you have a protocol for vacuum processing of blood samples with the QIAamp DNA Blood Midi or Maxi Kits?
Yes, please follow the Supplementary Protocol 'Purification of DNA from whole blood using the QIAamp DNA Blood Midi or Maxi Kit and vacuum processing on the QIAvac 24 Plus' (QA33).  Please contact Technical Service for this protocol.
FAQ ID -1011
Do any of the kit components or the product packaging of the QIAamp 96 DNA Blood Kit contain latex?
There is no latex in either the product or the packaging of the QIAamp 96 DNA Blood Kit.
FAQ ID -318
Is it possible to isolate DNA from bone marrow with the QIAamp DNA Blood Kits?

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 10and 1 x 108, respectively.

FAQ ID -563
Do you have a protocol for isolation of genomic DNA from saliva and mouthwash?

Yes, we have the following protocols:

  • Isolation of genomic DNA from saliva and mouthwash using the QIAamp DNA Mini Kit; vacuum procedure (QA19v)
  • Isolation of genomic DNA from saliva and mouthwash using the QIAamp DNA Mini Kit; spin procedure (QA19s)
  • Isolation of genomic DNA from saliva using the DNeasy Blood & Tissue Kit; spin procedure (DY07)
FAQ ID -917
When should carrier be used with the QIAamp DNA Mini or the DNeasy Blood & Tissue Kit?

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.

FAQ ID -100
How can I precipitate genomic DNA using isopropanol?

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.

 

Procedure

  1. Adjust the salt concentration, for example, with sodium acetate (0.3 M, pH 5.2, final concentration) or ammonium acetate (2.0–2.5 M, final concentration).
  2. Add 0.6–0.7 volumes of room-temperature isopropanol to the DNA solution and mix well.
  3. Centrifuge the sample immediately at 10,000–15,000 x g for 15–30 min at 4°C
  4. Carefully decant the supernatant without disturbing the pellet.
  5. Wash the DNA pellet by adding 1–10 ml (depending on the size of the preparation) of room-temperature 70% ethanol. This removes co-precipitated salt and replaces the isopropanol with the more volatile ethanol, making the DNA easier to redissolve.
  6. Centrifuge at 10,000–15,000 x g for 5–15 min at 4°C.
  7. Carefully decant the supernatant without disturbing the pellet.
  8. Air-dry the pellet for 5–20 min (depending on the size of the pellet).
  9. Redissolve the DNA in a suitable buffer.

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.

FAQ ID -2953
Do you have a protocol for isolation of genomic DNA from dried blood spots using the QIAamp 96 DNA Blood Kit?

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.

FAQ ID -919
How do I perform a DNA precipitation to concentrate my sample?
  • Add 1/10 volume of 3 M Na-Acetate pH 5.2, and 2 to 2.5 volumes of ice-cold 100% ethanol to the DNA sample
  • Mix, and store at –20°C for at least 1 h to precipitate the DNA
  • Recover the precipitated DNA by centrifugation at full speed in a microcentrifuge for 15–20 min
  • Pour off the ethanol and wash the pellet twice with room-temperature 70% ethanol
  • Allow the DNA pellet to air-dry
  • resuspend the DNA in a suitable volume of sterile TE buffer or distilled water
FAQ ID -305
Do you have a protocol for isolation of bacterial DNA from soil?
FAQ ID -923
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.

FAQ ID -761
Do you have a protocol for detection of HBV DNA by PCR?

Yes, please follow the User-Developed Protocol 'Detection of HBV DNA by PCR' (QA15). The procedure is for use with the QIAamp DNA Blood Mini Kit.  Please contact Technical Service for this protocol.

FAQ ID -915
Can I use your QIAvac 24 Plus for extraction of DNA from tissues using the DNeasy Blood & Tissue Kit?

We do not recommend using a vacuum manifold for DNA extraction of tissues using the QIAamp Blood & Tissue Kit. Viscosity of the lysates varies from sample to sample; therefore, the DNeasy mini spin columns are at great risk of clogging.

FAQ ID -9155
Is the quality and size of DNA extracted with the QIAamp DNA Mini and QIAamp Blood Mini kit good enough to generate DNA-libraries for next generation sequencing (NGS)?
The size of DNA obtained with QIAamp DNA Mini and QIAamp Blood Mini kit ranges between 100 bp and 50 kb, with 30 kb fragments predominating. The 30 kb fragments are a good starting point for most of the library preparations. Furthermore, the purified DNA is free of protein, nucleases, and other contaminants and inhibitors, and therefore is suitable for NGS.
FAQ ID - 3518
Do you have stability data for genomic DNA isolated with the QIAamp DNA Blood Mini Kit?

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.   

FAQ ID -518
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
What is the minimum number of cells that can be efficiently processed with the QIAamp DNA Mini and QIAamp Blood Mini kits and what is the expected yield?
In QIAGEN labs, we have isolated DNA from a minimum of 1000 cells or 1 ul of whole blood sample. The expected yield from 1 ul of whole blood with 4000-7000 leukocytes is 30 ng of genomic DNA. The expected yield of genomic DNA from a single eukaryotic cell is 6 pg. However, please bear in mind that for these small quantities, we would recommend the QIAamp DNA Micro kit instead.
FAQ ID - 3516
Are QIAamp DNA isolation kits suitable for apoptosis studies?

Yes. We have used the QIAamp DNA Blood Mini Kit to purify DNA fragments as small as 168 base pairs. Our product profile for this kit shows a picture of the apoptotic banding pattern obtained after storage of blood samples at 4°C for extended periods of time prior to isolating DNA.

 

 

FAQ ID -149
What dedicated QIAcube Kits are available?
What is the composition of buffer AE?

The composition of Buffer AE is:

  • 10 mM Tris-Cl
  • 0.5 mM EDTA; pH 9.0.
FAQ ID -730
What is the average amount of DNA and RNA present in 1 ml normal serum?

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.

FAQ ID -635
Why does the QIAamp DNA Mini Tissue Protocol require both ATL and AL buffer, while the Blood Protocol only uses AL?

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.

FAQ ID -633
How can QIAGEN Protease and Proteinase K be inactivated?

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.

FAQ ID -315
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
Do I need to have EDTA in the buffer in which I am going to store my isolated genomic DNA?

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.

FAQ ID -754
Do you have a protocol for Streptokinase treatment of clotted blood?
Yes, please follow the User-Developed Protocol 'Streptokinase treatment of clotted blood' (QA12). The protocol is for use with the QIAamp DNA Blood Mini Kit .
FAQ ID -912