cador Pathogen 96 QIAcube HT Kit

The cador Pathogen 96 QIAcube HT Kit provides fast and reliable automated purification of viral RNA and DNA and bacterial DNA from a range of animal samples — including whole blood, serum, plasma, body fluids, swabs, washes, tissue, and feces. High-quality nucleic acids are ready for use in a variety of sensitive downstream applications.

Purification of nucleic acids from undiluted animal whole blood is quick and simple, with minimal risk of silica membrane clogging. Automated processing of samples in 96-well format is precise and repeatable between different purification runs performed on different days, providing not only convenience but also peace of mind (see figure Highly repeatable viral RNA isolation).

The QIAcube HT platform with the cador Pathogen QIAcube HT Kit allows parallel purification of different nucleic acids from a range of sample types using just one universal instrument protocol. Co-purification of viral RNA and bacterial DNA is simple and reliable and provides the same high-quality nucleic acids as manual processing with the QIAamp cador Pathogen Mini Kit (see figure Co-isolation of viral RNA and bacterial DNA). Compared with a kit from another supplier, the cador Pathogen 96 QIAcube HT Kit displays high performance, and succeeds even when the other kit falls short (see figure High performance protocol).

The cador Pathogen 96 QIAcube HT Kit enables automated purification of pathogen nucleic acids — viral RNA and DNA and bacterial DNA — from up to 200 μl of animal whole blood, serum, plasma, swabs, or washes using the QIAcube HT instrument. By applying the pretreatments described in the cador Pathogen 96 QIAcube HT Handbook, tissue and fecal samples can also be processed. The procedure yields high-quality DNA and RNA that perform well in PCR, RT-PCR, and other downstream applications.

The cador Pathogen 96 QIAcube HT Kit combines the selective binding properties of a silica-based membrane with a high-throughput 96-well format, and is designed for fully automated, simultaneous processing of 24–96 samples on the QIAcube HT instrument.
 

Kit specifications

* Time for the standard protocol, excluding any sample pretreatments.

Specification	Description
Number of samples	24–96 samples (to be processed in increments of 8)
Sample input volume	Up to 200 μl
Elution volume	150 μl
Duration	96 samples in approximately 90 minutes* 24 samples in approximately 60 minutes* Each additional 8 samples extend the processing time by 5 minutes

Samples are lysed under highly denaturing conditions at room temperature in the presence of proteinase K and Buffer VXL, which together ensure the inactivation of nucleases. Adding Buffer ACB adjusts the binding conditions for the co-purification of DNA and RNA. The lysate is then transferred to a QIAamp 96 plate. As vacuum is applied, DNA selectively binds to the QIAamp membrane as contaminants pass through. Three efficient wash steps remove the remaining contaminants and enzyme inhibitors, and nucleic acids are eluted in Buffer AVE.

Most common fluid samples can be processed directly using the main protocol, which involves the lysis, binding, wash, and elution steps. However, depending on the starting material and the target pathogen, one of the pretreatment protocols may be needed (see the table "Pretreatments for the cador Pathogen 96 QIAcube HT Kit protocol"). After pretreatment, the samples undergo the same procedure as for fluid samples, enabling automated parallel processing on the QIAcube HT.

Pretreatments for the cador Pathogen 96 QIAcube HT Kit protocol

* 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.

Name	Application
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 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 F1	Isolation of viral RNA and DNA from fecal samples
Pretreatment F2	Isolation of bacterial and viral DNA from fecal samples

The cador Pathogen 96 QIAcube HT 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