QIAamp DSP Circulating NA Kit

For concentration and purification of free-circulating DNA and RNA from human plasma samples

Features

  • Concentrated nucleic acids; high input, low elution volumes
  • Efficient recovery of DNA and RNA
  • No organic extraction or ethanol precipitation
  • Removal of contaminants and inhibitors
  • Produced under GMP manufacturing conditions
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QIAamp DSP Circulating NA Kit

Cat. No. / ID: 61504

For 50 preps: includes QIAamp Mini Columns, Buffers, Carrier RNA, QIAGEN Proteinase K, and Tubes.
$1,309.00
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The QIAamp DSP Circulating NA Kit is intended for in vitro diagnostic use.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Product Details

The QIAamp DSP Circulating NA Kit is for concentration and purification of free-circulating DNA and RNA from plasma for in vitro diagnostic use.

Performance

The QIAamp DSP Circulating NA Kit provides ease-of-use in diagnostic workflows, and is manufactured in accordance with GMP requirements. The procedure isolates and purifies high quality nucleic acids for use in downstream applications. The concentration of free-circulating nucleic acids in human blood plasma is usually low and varies considerably between individuals, ranging from 1–100 ng/ml in human samples. Nucleic acids are efficiently purified and concentrated from starting materials that contain low concentrations of mostly fragmented DNA and RNA (typically 1–100 ng/ml circulating DNA in human plasma).

Principle

The QIAamp DSP Circulating NA Kit simplifies isolation of circulating DNA and RNA from plasma samples by omitting phenol-chloroform extractions. Nucleic Acids bind specifically to the QIAamp Mini columns and contaminants pass through. PCR inhibitors are completely removed in the wash steps, leaving pure nucleic acids for elution with the provided buffer.

The QIAamp DSP Circulating NA Kit combines the selective binding properties of silica-based membrane with flexible elution volumes between 20 µl and 150 µl.

Tube extenders and vacuum processing on the QIAvac 24 Plus enable starting sample volumes of up to 5 ml (figure  Tube extenders for processing on the QIAvac 24 Plus). Flexible elution volumes between 20 µl and 150 µl allow concentration of nucleic acid species that are present in low concentrations.

See figures

Procedure

The QIAamp DSP Circulating NA 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. Nucleic acids are efficiently purified and concentrated (figure  QIAamp DSP Circulating NA Kit procedure). Additionally, the QIAamp DSP Circulating NA Kit offers two distinct protocols: the Classic and Breeze protocols (figure  Comparison of Classic and Breeze workflows). The Classic protocol is for processing up to 5 ml plasma in 1 ml steps, and constitutes the unchanged protocol provided in the QIAamp DSP Circulating NA Kit Handbook, Revision 3 (R3). The Breeze protocol is for processing up to 5 ml plasma in 1 ml steps, but has been optimized for low hands-on and turnaround time.

See figures

Applications

ccfDNA purified using the  QIAamp DSP Circulating NA Kit is ready for use in a wide range of downstream applications, including NGS, RT-PCR, digital PCR, multiplex PCR and quantitative PCR.

Supporting data and figures

Publications

Implementing prenatal diagnosis based on cell-free fetal DNA: accurate identification of factors affecting fetal DNA yield.
Barrett AN; Zimmermann BG; Wang D; Holloway A; Chitty LS;
PLoS One; 2011; 6 (10):e25202 2011 Oct 4 PMID:21998643

FAQ

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 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