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Каталог

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QIAamp cador Pathogen Mini Kit

For isolation of viral RNA and DNA and bacterial DNA from a variety of animal sample types

  • Same protocol for viral RNA, viral DNA, and bacterial DNA
  • Suitable for many samples, including whole blood, serum, and tissue
  • Even suitable for viral RNA in undiluted animal whole blood
  • Efficient removal of inhibitors and contaminants
  • Isolated nucleic acids ready for analysis by real-time PCR or RT-PCR

Effective April 1, 2018, product available exclusively from INDICAL BIOSCIENCE (formerly QIAGEN Leipzig).

The QIAamp cador Pathogen Mini Kit simplifies the isolation of viral RNA and DNA and bacterial DNA from a whole range of animal samples, including undiluted whole blood, serum, swabs, and tissue. Using a single, rapid spin-column procedure, contaminants and inhibitors are removed to yield pathogen nucleic acids that are ready for use in downstream applications, such as real-time PCR and RT-PCR. The procedure can be fully automated on the QIAcube.

Артикул: 54106
QIAamp cador Pathogen Mini Kit (250)
For 250 RNA/DNA preps: 250 QIAamp Mini Spin Columns, Carrier RNA, Proteinase K, Collection Tubes (2 ml), RNase-free buffers
Effective April 1, 2018, product available exclusively from INDICAL
Артикул: 54104
QIAamp cador Pathogen Mini Kit (50)
For 50 RNA/DNA preps: 50 QIAamp Mini Spin Columns, Carrier RNA, Proteinase K, Collection Tubes (2 ml), RNase-free buffers
Effective April 1, 2018, product available exclusively from INDICAL


Performance

Data show that high and identical purification efficiency can be achieved with both the manual and automated protocols for viral RNA isolation. Bovine serum was spiked with virus particles from cell culture. The sample was serially diluted 1:10 with bovine serum and processed in duplicate using the QIAamp cador Pathogen Mini Kit either manually or using the automated protocol on the QIAcube. Viral RNA was detected using the QuantiFast Pathogen RT-PCR +IC Kit with target-specific primers and probe.

The robustness of the protocol is demonstrated by the high intra- and inter-assay repeatability. Whole blood samples from different animals were spiked with virus particles from cell culture and then frozen at -20°C. Three replicates for each sample were processed on three different days with the QIAamp cador Pathogen Mini Kit. The viral RNA was amplified with the QuantiFast Pathogen RT-PCR +IC Kit using target-specific primers and probe.

The kit also successfully isolates bacterial DNA from animal samples. Different animal samples were spiked with Gram-negative or Gram-positive bacteria and pretreated as necessary (see the table "Pretreatments for the QIAamp cador Pathogen Mini Kit protocol"), and then the automated isolation protocol was run on the QIAcube. The bacterial DNA was successfully purified. PCR setup with parallel preparation of reaction mixes for both bacterial targets was performed using the QIAgility, and the bacterial DNA was amplified with the QuantiFast Pathogen +IC Kit with target-specific primers and probes.

Principle

The QIAamp cador Pathogen Mini Kit combines the features of several sample preparation kits based on spin-column technology to permit the purification of viral RNA and DNA and bacterial DNA from animal fluid and tissue samples. The spin-column procedure uses optimized buffers and enzymes to lyse samples and stabilize the pathogen nucleic acids. This RNA and/or DNA binds to the QIAamp silica membrane, while contaminants pass through. Wash buffers are used to completely remove PCR inhibitors, such as divalent cations and proteins, and the pure pathogen nucleic acids are then eluted in Buffer AVE.

QIAamp cador Pathogen Kit technology yields pathogen RNA and DNA from animal samples that is ready for use in downstream applications such as real-time PCR and RT-PCR.

In contrast to other kits based on silica membranes, up to 200 µl of undiluted animal whole blood samples can be processed with the QIAamp cador Pathogen Mini Kit without clogging the filter. See the table "Key features of the QIAamp cador Pathogen Mini Kit".

Key features of the QIAamp cador Pathogen Mini Kit

Format and processing Mini spin columns, manual centrifuge protocol, or automated centrifuge protocol using the QIAcube
Target nucleic acids Viral RNA, viral DNA, and bacterial DNA
Sample source types Animal whole blood, serum, plasma, other fluids, swabs, washes, and tissues
Sample size Up to 200 ?l or 25 mg
Preparation time 20 to 40 minutes
Elution volume 50 to 150 ?l

Procedure

Most common fluid samples can directly be processed using the main protocol, which involves the lysis, binding, wash, and elution steps described in the "Principle" section.

Tissue samples and samples containing difficult-to-lyse bacteria require the appropriate pretreatment (see the table "Pretreatments for the QIAamp cador Pathogen Mini Kit protocol"). After pretreatment, the samples undergo the same procedure as for fluid samples, enabling parallel processing either manually or in an automated procedure on the QIAcube.

Pretreatments for the QIAamp cador Pathogen Mini Kit protocol

Name Application
Pretreatment B1 For difficult-to-lyse bacteria* in whole blood or pretreated tissue
Pretreatment B1 For difficult-to-lyse bacteria* in whole blood or pretreated tissue
Pretreatment B2 For difficult-to-lyse bacteria* in cell-free fluids
Pretreatment B3 For easy-to-lyse bacteria in high-volume cell-free fluids
Pretreatment T1 Mechanical disruption of tissue for extraction of viral RNA and DNA
Pretreatment T2 Enzymatic digestion of tissue for extraction of viral and bacterial DNA
Pretreatment T3 Rapid partial disruption of tissue for extraction of viral RNA and DNA and bacterial DNA
Pretreatment T4 Organic extraction for extraction of viral RNA and DNA and DNA from easy-to-lyse bacteria* from difficult tissue
* Gram-positive bacteria are difficult to lyse due to their rigid cell wall. However, some Gram-negative bacteria are also difficult to lyse, and benefit from pretreatment B1 or B2.

Applications

The QIAamp cador Pathogen Mini Kit yields pure, high-quality viral RNA, viral DNA, and bacterial DNA that is suitable for a wide range of downstream applications, including real-time PCR and RT-PCR for:

  • Animal pathogen identification
  • Animal pathogen genotyping
  • Epidemiology
  • Infectious disease research