QIAamp Circulating Nucleic Acid Kit

For isolation of free-circulating DNA and RNA from human plasma or serum

  • Concentration of nucleic acids, with high input and low elution volumes
  • Efficient recovery of fragmented DNA and RNA
  • No organic extraction or ethanol precipitation
  • Removal of contaminants and inhibitors

The QIAamp Circulating Nucleic Acid Kit greatly simplifies concentration and purification of free-circulating DNA and RNA from plasma or serum. The procedure can be fully automated on the QIAcube.

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QIAamp Circulating Nucleic Acid Kit (50)
For 50 preps: QIAamp Mini Columns, Tube Extenders (20 ml), QIAGEN Proteinase K, Carrier RNA, Buffers, VacConnectors, and Collection Tubes (1.5 ml and 2 ml)
55114
938,00 €
The QIAamp Circulating Nucleic Acid Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Reproducible recovery of circulating DNA.|Improved recovery of fragmented DNA.|Efficient purification of circulating miRNA.|Efficient recovery of methylated DNA.|
DNA fragments (200 bp and 1000 bp) in equal amounts were added to 24 plasma samples. DNA was purified from 5 ml plasma using the QIAamp Circulating Nucleic Acid Kit, with an elution volume of 80 μl. DNA yield was quantified by duplex, real-time PCR of 66 bp targets specific for each DNA fragment using the QuantiTect Multiplex PCR Kit.|Circulating DNA was purified from 5 ml plasma using the QIAamp Circulating Nucleic Acid Kit or from 1 ml plasma using the QIAamp MinElute Virus Vacuum Kit, with elution volumes of 100 μl. DNA yield was quantified by duplex, real-time PCR of a 66 bp amplicon and a 500 bp amplicon within the 18S rRNA gene using the QuantTect Multiplex PCR Kit. The difference between amplification of the 500 bp amplicon and 66 bp amplicon shows that the DNA is fragmented, resulting in a lower abundance of intact 500 bp target sequences compared with the shorter 66 bp target. (Note that the difference is greater than the fivefold difference in sample input volumes.)|Circulating RNA was purified from 4 ml pooled plasma using the QIAamp Circulating Nucleic Acid Kit, with an elution volume of 50 μl. The standard kit protocol was compared with the specialized protocol for miRNA. miRNA 30b yield was quantified using a TaqMan microRNA assay (Applied Biosystems), and miRNA 16 yield was quantified using a Human miScript Assay (QIAGEN). The lower CT values with the miRNA protocol indicate higher yields of the miRNA species.|Spiked methlyated DNA and non-target poly-A RNA were purified from 4 ml plasma using the QIAamp Circulating Nucleic Acid Kit, with an elution volume of 45 μl. Bisulfite conversion was carried out using the EpiTect Bisulfite Kit (QIAGEN), and methylation-specific PCR (MSP) was performed using real-time MSP assays and components of the QuantiTect Multiplex PCR Kit on the ABI PRISM 7900HT Sequence Detection System. Data from two fully methylated loci were analyzed.|
Performance

Analysis of tumor-specific extracellular DNA fragments and mRNAs in the blood can enable specific detection of tumor types from a simple blood sample. These circulating nucleic acids are present in serum or plasma usually as short fragments, <1000 bp (DNA) or <1000 nt (RNA). In addition, miRNAs, as small as 21 nt, have the potential to provide biomarkers for certain cancers and disease states.

The QIAamp Circulating Nucleic Acid Kit enables efficient purification of these circulating nucleic acids from human plasma or serum and other cell-free body fluids. Efficient purification with reproducible yields provides a representative population of the circulating nucleic acids in blood (see figure "Reproducible recovery of circulating DNA"). Tube extenders and vacuum processing on the QIAvac 24 Plus enable starting sample volumes of up to 5 ml, and flexible elution volumes between 20 μl and 150 μl allow concentration of nucleic acid species that are present in low concentrations. The kit provides advanced technology of selective binding to a silica-based membrane for improved recovery of fragmented nucleic acids (see figure "Improved recovery of fragmented DNA").

Purification of circulating RNA, without copurification of DNA, is possible with DNA digestion using the RNase-Free DNase Set. A specialized protocol provides highly efficient purification of small RNA, such as miRNAs (see figure "Efficient purification of circulating miRNA"). Methylated DNA can be efficiently purified using the QIAamp Circulating Nucleic Acid Kit. The purified DNA maintains its methylation, allowing bisulfite conversion for analysis of the methylation state (see figure "Efficient recovery of methylated DNA").

The purified and concentrated circulating DNA and RNA is free of proteins, nucleases, and other impurities, and is ready to use in wide range of downstream applications, including:

PCR and quantitative real-time PCR and RT-PCR
Biomarker research and validation for blood-based cancer detection
Viral nucleic acid detection 
Principle

The QIAamp Circulating Nucleic Acid Kit simplifies isolation of circulating DNA and RNA from plasma or serum. No phenol–chloroform extraction is required. Nucleic acids bind specifically to the QIAamp Mini column, while contaminants pass through. PCR inhibitors, such as divalent cations and proteins, are completely removed in 3 wash steps, leaving pure nucleic acids to be eluted in a buffer provided with the kit. The QIAamp Circulating Nucleic Acid technology yields circulating DNA and RNA from human plasma, serum, or urine.

The QIAamp Circulating Nucleic Acid Kit combines the selective binding properties of silica-based membrane with flexible elution volumes between 20 and 150 μl. Circulating RNA can be purified with DNA digestion using the RNase-Free DNase Set.

Procedure

The QIAamp Circulating Nucleic Acid procedure includes 4 steps (lyse, bind, wash, and elute) that are carried out using QIAamp Mini columns on a vacuum manifold. The simple procedure is highly suited for simultaneous processing of multiple samples, providing nucleic acids in less than 2 hours per 24 samples. If the QIAamp Circulating Nucleic Acid Kit is used for isolation of viral RNA and DNA, the performance cannot be guaranteed for every virus species and must be validated by the user.

Applications

The QIAamp Circulating Nucleic Acid Kit efficiently purifies and concentrates free-circulating DNA, RNA, miRNA, and viral nucleic acids from starting materials that contain low concentrations of mostly fragmented DNA and RNA (typically 1–100 ng/ml circulating DNA in human plasma). Starting sample volumes can be up to 5 ml.

The QIAamp Circulating Nucleic Acid Kit purifies and concentrates nucleic acids from the following sample types:

Human plasma
Serum
Urine
Feature
Specifications
Applications PCR, real-time PCR
CE/FDA/IVD compatible -
Elution volume 20–150 µl
Format QIAamp Mini Columns
Main sample type Serum, plasma
Processing Manual (vacuum)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Free-circulating DNA, RNA and miRNA, viral DNA, viral RNA
Sample amount 1-5 ml
Technology Silica technology
Time per run or per prep <2 hour for 24 preps
Yield Varies, due to donor-to-donor variations and disease status

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Kit Handbooks
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For concentration and purification of free-circulating DNA and RNA from human plasma or serum
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