QIAsymphony SP/AS instruments

The following disinfectants and detergents are recommended for cleaning the QIAsymphony SP:

• Mikrozid® Tissues (Schülke & Mayr GmbH) — 150 tissues with 235 g agent (consists of 24g (94%) ethanol and 35g 1-propanol per 100g agent)
• Mikrozid Liquid (Schülke & Mayr GmbH) — ethanol-based disinfectant for spraying onto surfaces (consists of 25 g ethanol and 35 g 1-propanol per 100 g agent solution)
• RBS®-35 Detergent Concentrate (Pierce Chemical Co.) — anionic and nonionic surfactant based detergent for cleaning the touch screen and worktable door.
• RNaseZap® (Ambion) — for complete removal of RNase contamination from glass and plastic surfaces
• 0.1 M NaOH — for cleaning the worktable and racks to remove RNase contamination (alternative to RNaseZap)

Europe

• Lysetol® AF (Gigsept® Instru AF) (Schülke & Mayr GmbH) — disinfectant for submerging worktable items (consists of 14 g cocospropylene-diamine-guanidine diacetate, 35 g phenoxypropanols, and 2.5 g benzalkonium chloride per 100 g Lysetol AF, with anticorrosion components, fragrance, and 15-30% nonionic surfactants

USA

• DECON-QUAT® 100 (Veltek Associates, Inc.) — quaternary ammonium salt based disinfectant concentrate for submerging worktable items (contains 5% alkyldimethylbenzylammonium chloride and 5% alkyldimethylethylbenzylammonium chloride)

Note: If you want to use disinfectants different from those recommended, ensure that their compositions are similar to those described above.

No. Modified QIAGEN protocols or custom protocols for QIAsymphony SP can be developed to meet your specific requirements and purchased from QIAGEN.

No. The use of internal controls with the QIAsymphony Virus/Bacteria Kits is not mandatory. When using internal controls, they are added to the carrier RNA mixture (total volume of the mixture must be 120 µl).

Yes. Any number of samples between 1 and 96 can be processed on the QIAsymphony SP.

The barcode on the adapter at the cooling elution position is read by default. This can be changed in the configuration settings which are available from the tools menu if you have logged in as Supervisor. The option is on the tab Process SP1 and is called ”On which elution slots should the bar codes be scanned”. To disable the barcode reading, put the setting to ”none”.

For purification of RNA from blood on the QIAsymphony SP, we recommend using the QIAsymphony PAXgene RNA Kit, since it includes stabilization of gene expression profiles.

The QIAsymphony Virus/Bacteria Mini Kit is used with the 'Cellfree_200' and 'Complex_200' protocols.

The QIAsymphony Virus/Bacteria Midi Kit is used with the 'Cellfree_500’, 'Cellfree_1000', ‘Complex_400’ and 'Complex_800' protocols.

Part number for O-rings for pipetting channels of QIAsymphony SP and AS instruments is 9019168. There are 32 O-rings included in one bag.

You will also require the tool to change the O-rings. Part number for this tool is: 9019164. This tool set includes one bag of 32 O-rings.

We recommend replacing O-rings on all pipetting channels, once a month.

No. All solid materials (e.g., swabs, blood card punches, chewing gum) must be removed completely from the lysate before performing automated extraction on the
QIAsymphony SP. The QIAshredder can be used to increase sample recovery from some solid materials. For more information, see the corresponding pretreatment protocol.

For purification of RNA on the QIAsymphony SP, tissue samples require mechanical disruption and homogenization in the supplied lysis buffer (Buffer RLT Plus), e.g., by using the TissueLyser or TissueRuptor. Cultured cells should be lysed by vortexing in the lysis buffer contained in the kit.

Please navigate to Tools/Instrument Report/Create + Save to USB.

No. We do not recommend use of carrier RNA with the QIAsymphony Virus Blood 200 Protocol.

In general, magnetic particles are not carried over into QIAsymphony eluates. However, if high amounts of genomic DNA are purified (large number of white blood cells in the sample) magnetic-particle carryover may be observed. If carryover does occur, magnetic particles in eluates will not affect most downstream applications. If magnetic particles need to be removed before performing downstream applications, tubes or plates containing eluates should first be placed in a suitable magnet and the eluates transferred to a clean tube.

Tissue: If no information about the expected yield is available, we recommend starting with 25 mg sample material. Depending on the yield obtained, the sample size can be increased in subsequent preparations to a maximum of 50 mg.

FFPE: Up to 4 sections, each with a thickness of up to 10 µm, or 8 sections with a thickness of up to 5 µm and a surface area of up to 250 mm2, can be combined in one preparation.

Eluates obtained on the QIAsymphony SP can be stored at 2–8°C for 3 days before use in downstream applications and should be placed at –20°C or –80°C for long-term storage.

Plasma, serum, and CSF samples can be processed using the Virus Cellfree protocols of the QIAsymphony Virus/Bacteria Kits. Respiratory samples (sputum, BAL, aspirates, dried swabs, transport media), and urogenital samples (urine, urogenital swabs, transport media) can be processed using the Pathogen Complex protocols.

DNA yields strongly depend on the type and source of the tissue processed on the QIAsymphony SP. Approximately 70–80 µg DNA can be isolated from 25 mg of spleen, 40–50 µg from 25 mg liver, and 10–15 µg from 50 mg muscle.

The maximum amount of tissue that can be processed on the QIAsymphony SP using QIAsymphony DNA Mini Kit is 50 mg, and the maximum number of cultured cells is 1 x 10⁷.

See details for upper limits of various starting materials in the table below:

The QIAsymphony system has an integrated UV lamp. UV decontamination should be performed daily. It helps to reduce possible pathogen contamination of the QIAsymphony SP/AS worktables. The efficiency of inactivation has to be determined for each specific organism and depends, for example, on layer thickness and sample type. QIAGEN cannot guarantee complete eradication of specific pathogens.

A total of 192 samples can be processed with one QIAsymphony DNA Mini Kit and the Virus Blood 200 protocol (96 samples per reagent cartridge).

QIAGEN protocols on the QIAsymphony SP can be stopped if there is an emergency by pressing "Pause". To confirm that you want to stop the protocol run, press "Stop". To cancel the stop protocol command, press "Continue".

Note: If a protocol run is stopped, the run cannot be restarted. Samples must be processed manually. For more information, refer to the QIAsymphony User Manual.

Yes, the QIAsymphony RNA Kit is compatible with stabilized samples. We recommend stabilizing tissue samples with RNAprotect Tissue Reagent or AllProtect Tissue Reagent, or by flash-freezing. Cultured cells can be lysed directly in cell culture dishes, or after pelleting. Alternatively, cells can be stabilized using RNAprotect Cell Reagent.

Sample volumes of 200 µl can be processed with the Virus Blood 200 Protocol using the QIAsymphony DNA Mini Kit.

Yes. Samples have to be completely lysed and homogenized prior to automated processing on the QIAsymphony SP platform using QIAsymphony DNA Kits.

Samples are lysed by Proteinase K digestion in lysis buffers optimized for tissue, cultured cells, and bacteria, respectively. For lysis of gram-positive bacteria, lysozyme is used in addition to Proteinase K. Lysed samples are homogenized by pipetting up and down, and insoluble material is removed by centrifugation (see the QIAsymphony DNA Handbook for details).

Samples should be processed on the QIAsymphony within 24 hours of collection. Samples should be transported and stored at 2–25°C.

No. The specified lysate volumes are optimal for QIAsymphony DNA Investigator Kit protocols. Lower volumes (150 µL,400 µL, 900 µL, respectively) can be used.

However, to maximize yields, we recommend transferring as much of the lysate as possible.
The QIAshredder can be used to increase recovery from some sample types. For more information, see the corresponding pretreatment protocol.

The QIAsymphony SP instrument has the following weight and dimensions:

• Weight: 178 kg (392.4 lb.); 220 kg (485 lb.) with packaging
• Width: 130 cm (51.2 in.)
• Depth: 75 cm (29.5 in.); 125 cm (49.2 in.) with drawer open
• Height (door closed): 103 cm (40.6 in.)
• Height (door open): 163 cm (64.2 in.)

Yes, QIAsymphony SP can be connected to a lab information management system. Result files can be exported in html format. At a later date, users will be able to import sample worklists to the QIAsymphony SP.

The QIAsymphony RNA Kit enables purification of RNA from cultured cells and easy-to-lyse tissues, such as kidney, liver, and spleen on the QIAsymphony SP. A large-volume protocol is available that enables processing of larger sample amounts.

• Cultured cells: up to 3x10⁶ cells (standard protocol); up to 1x10⁷ (large volume protocol)
• Tissues: up to 20 mg of most tissues (standard protocol); up to 50 mg (large-volume protocol)

For particularly DNA-rich tissues, such as spleen or thymus, we recommend either using no more than half the maximum amount, or using DNase at a higher concentration (see the QIAsymphony RNA Handbook; additional DNase required).

Sample volumes that can be processed with the QIAsymphony Virus/Bacteria Kits depend on the protocol:

• 'Cellfree_200' and 'Complex_200':  200 µl
• 'Complex_800':                            800 µl
• 'Cellfree_1000':                           1000 µl

Human whole blood samples treated with EDTA or citrate can be processed using the QIAsymphony Virus Blood 200 protocol with the QIAsymphony DNA Mini Kit. We do not recommend using heparin-treated blood samples.

Yes. The DNA Tissue protocols on the QIAsymphony SP purify genomic and mitochondrial DNA.

You can find this information in the respective labware list. All tubes which require an insert in the sample input carrier or plates are not suitable.

Yes. QIAsymphony DNA Investigator Kit protocols purify both genomic and mitochondrial DNA.

Strongly denaturing conditions in the lysis buffer (Buffer RLT Plus) of the QIAsymphony RNA Kit keep RNases in an inactive state. Keeping homogenates on the QIAsymphony instrument at room temperature (20-25°C) for over 6 hours does not impair RNA integrity.

Final elution volumes of 60 µl, 85 µl, 110 µl, or 165 µl can be selected using the QIAsymphony Virus Blood 200 protocol. Please note that the initial elution volumes are 95 µl, 120 µl, 145 µl, or 200 µl. When calculating the amount of internal control(s) as well as the titer of the processed sample, it is necessary to take into consideration the initial volume of elution buffer that is used for each sample.

No. Protocols on the QIAsymphony SP cannot be modified by users. However, protocols allow elution volumes to be selected within a predefined range. Customized protocols for QIAsymphony SP will be available on request.

Code 39, code 128 and subtypes, Codabar

The High Content protocol for DNA extractions on the QIAsymphony SP is optimized for samples with an expected DNA yield of more than 30 µg. Up to 90 µg DNA can be purified from tissues with high cell densities (e.g., spleen) or large numbers of cultured cells.

Yes. At the end of each protocol run on the QIAsymphony SP, a report file is generated that contains important information such as number of samples, liquid volumes measured during the load check, and run time. A log file is also generated that contains information about instrument actions, such as movement of the robotic arm.

Depending on the application, elution volumes on the QIAsymphony SP range from 30 µl to 500 µl.

The large-volume protocol of the QIAsymphony DNA Investigator Kit is intended for samples that can not be submerged completely during lysis in volumes less than 1 ml, as well as samples that are very absorbent.

Please see expected yields of total RNA isolated with the QIAsymphony RNA Kit on the QIAsymphony SP in the table below:

*RNA yields strongly depend on the type and quality of the sample material.

Various types of tissue, cultured cells, and bacterial pellets can be processed using the tissue protocols for the QIAsymphony DNA Kits on the QIAsymphony SP.

Pre-treatment of samples used with the QIAsymphony Virus/Bacteria Kits can improve nucleic acid extraction for some sample types. Viscous samples (e.g., sputum, respiratory aspirates) might require liquefaction to enable pipetting. Pretreatment protocols are provided in the QIAsymphony Virus/Bacteria Handbook.

Yes. Magnetic particles used in the QIAsymphony SP System will bind RNA and total DNA. If isolation of RNA-free DNA is required, RNase A must be added to the lysate before isolation of DNA.

Power consumption of a QIAsymphony SP instrument is 800 VA. Therefore, the approximate thermal output is 2729 BTU/h.

If the QIAsymphony SP is connected to a QIAsymphony AS, then the power consumption is 1400 VA. The thermal output is ~4777 BTU/h.

QIAsymphony DNA Investigator Kit protocols are optimized for maximum sensitivity in extraction of typical forensic-casework and reference sample types, which usually have low DNA content. Due to optimized bead capacity, maximum DNA yields are approximately 3 µg.

Please navigate to the Product Page of the QIAsymphony Kit you are using; for example, the QIAsymphony RNA Kit, Cat No./ID: 931636.  Refer to the resources tab and download the labware document. The labware document will list the sample tubes that have been tested for your application. Please contact Technical Service if you wish to use sample tubes not listed in the labware document.

No. Carrier RNA is only used in casework protocols for the QIAsymphony DNA Investigator Kit to enable efficient extraction of minute quantities of DNA. Carrier RNA is not used in reference protocols.

No. The use of internal controls is not mandatory for use with the QIAsymphony Virus Blood 200 Protocol. If an internal control is not required, Buffer ATE without internal control is added to the samples. Buffer ATE must be placed in slot A of the sample drawer.

To ensure performance with the QIAsymphony DNA Investigator Kit, lysis volumes have been optimized according to sample type. In general, large or absorbent samples require larger buffer volumes than small and non-absorbent samples.

Buffer ATE of the QIAsymphony DNA Investigator Kit is a low-EDTA elution buffer optimized for long-term storage of DNA as well as performance in sensitive downstream applications. Alternatively, casework protocols using water for elution are available.

Buffer ATE contains a small amount of the preservative sodium azide. If the eluate is measured for absorbance the spectrophotometer should be blanked with the buffer ATE as sodium azide absorbs strongly at 230 nm. 

Each QIAsymphony RNA Kit enables preparation of up to 192 samples (96 per reagent cartridge) using the standard protocol on the QIAsymphony SP. If only the large-volume protocol is used, the number of samples that can be prepared per kit is reduced to 96 (48 per reagent cartridge). Any combination of standard and large-volume sample preparation can be performed.

The target nucleic acid for the Virus Blood 200 Protocol on the QIAsymphony is viral DNA from released as well as cell-associated viruses, co-purified with genomic DNA from blood cells.