QIAsymphony® DSP DNA Midi Kit Protocol Sheet

General Information

The QIAsymphony DSP DNA Kit is intended for in vitro diagnostic use. This protocol sheet is applicable for purifying total genomic and mitochondrial DNA from fresh or frozen human whole blood using the QIAsymphony SP and QIAsymphony DSP DNA Midi Kit.

Procedure Name: DNA_Blood_1000_V7_DSP Procedure

Default Control Set: ACS_Blood_1000_V7_DSP

Editable: Yes

Required Software Version: Version 4.0 or higher

Software Configuration for IVD Use: Default File 1

Sample Drawer

Sample Type: Human whole blood (EDTA, citrate, or heparin anticoagulated)

Sample Volume: Refer to the laboratory consumables list for specific sample tube types; available on the product page under the Resources tab: www.qiagen.com.

Primary Sample Tube: Refer to the laboratory consumables list; available on the product page under the Resources tab: www.qiagen.com.

Secondary Sample Tube: Refer to the laboratory consumables list; available on the product page under the Resources tab: www.qiagen.com.

Plugin: Refer to the laboratory consumables list; available on the product page under the Resources tab: www.qiagen.com.

Reagents and Consumables Drawer

Position A1 and/or A2: Reagent Kit (RC)

Position B1: Not applicable

Tip Rack Holder 1-17: Disposable filter tips, 200 µl or 1500 µl

Unit Box Holder 1-4: Unit boxes containing sample preparation reagents and 8-Rod Covers.

n/a: Not applicable.

Waste Drawer

Unit Box Holder 1-4: Empty unit boxes

Waste Bag Holder: Waste bags

Waste Bottle Holder: Empty waste bottles

Eluate Drawer

Elution Rack: Recommended to use Slot 1, cooling position. For more information, refer to the laboratory consumables list, available on the product page under the Resources tab: www.qiagen.com.

Plastic Consumables

* Fewer than 24 samples per batch reduces the number of disposable filter tips required per run.

** Each tip rack contains 32 filter tips.

The number of filter tips required includes filter tips for one-time scan for each reagent kit.

Each unit box contains 28 sample prep cartridges.

Each unit box contains 12 8-Rod Covers.

Note: The number of filter tips may differ from the number displayed on the touchscreen depending on the configuration. It is recommended to load the maximum possible number of tips.

Elution Volume

Select the elution volume on the touchscreen. Depending on the sample type and DNA content, the final eluate volume may be up to 15 µl less than the selected volume. As the elution volume may vary, it is recommended to use the automatic detection system to check the actual eluate volume, which does not verify the eluate volume before transfer. Lower elution volumes can increase the final DNA concentration, but may slightly decrease the yield. It is recommended to use an elution volume suitable for the intended downstream application.

Sample Preparation Material

Always wear appropriate laboratory coats, disposable gloves, and safety glasses when handling chemicals. For more information, refer to the relevant Safety Data Sheets (SDS), which can be obtained from the product supplier.

For general recommendations on collection, transport, and storage, refer to the approved CLSI guideline MM13-A, "Collection, Transport, and Storage of Specimens for Molecular Testing." Additionally, follow the manufacturer's instructions for the chosen sample collection device during sample preparation, storage, transport, and general handling.

Refer to the manufacturer's instructions for blood collection tubes for the automatic extraction of gDNA from peripheral whole blood according to ISO 20186-2:2019 (E).

Human Whole Blood

Fresh or frozen human whole blood samples anticoagulated with EDTA, citrate, or heparin can be used. If using fresh blood samples in primary tubes, mix thoroughly by inverting the tube several times before loading the sample onto the QIAsymphony SP. Frozen samples should be rapidly thawed in a 37°C water bath and gently mixed to ensure complete mixing, then allowed to equilibrate to room temperature (15–25°C) before starting the procedure. To ensure reliable sample transfer, avoid foaming in the sample tubes. Avoid blood clots in the sample; if necessary, transfer samples without clots to fresh tubes.

The yield and quality of purified DNA depend on the storage conditions of the blood. Fresher blood samples yield higher amounts. Short-term storage of up to 10 days is recommended at 2–8°C. For applications requiring maximum fragment size, such as Southern blotting, storage at 2–8°C for up to 3 days is recommended, as low-level DNA degradation may occur thereafter. Long-term storage (more than 10 days) should involve collecting blood in tubes containing standard anticoagulants (preferably EDTA if high molecular weight DNA is required) and storing at –20°C or –80°C.

Note: Sample stability is influenced by multiple factors and specific downstream applications. The information provided is based on typical downstream applications when using the QIAsymphony DSP DNA Midi Kit. Users are responsible for verifying the usage instructions for specific downstream applications in their laboratory and/or validating the overall workflow to establish appropriate storage conditions.

Eluate Storage

It is recommended to remove the eluate tubes from the "Eluate" drawer immediately after the run is completed. For overnight runs, the eluate tubes can remain in the QIAsymphony SP (up to 12 hours, including run time). Recommended environmental conditions are 18–26°C and 20–75% relative humidity. Depending on temperature and humidity, eluates may condense or evaporate.

Eluates can be stored short-term at room temperature for up to 2 weeks. Long-term storage is recommended at 2–8°C, –20°C, or –80°C. Frozen eluates should not be thawed more than three times.

Note: Eluate stability is influenced by multiple factors and specific downstream applications. The information provided is based on typical downstream applications when using the QIAsymphony DSP DNA Midi Kit. Users are responsible for verifying the usage instructions for specific downstream applications in their laboratory and/or validating the overall workflow to establish appropriate storage conditions.

Important Notes Before Starting

QIAsymphony magnetic beads simultaneously purify RNA present in the sample. To minimize the amount of RNA in the sample, add RNase A to the sample before starting the procedure. The final RNase A concentration should be 2 mg/ml.

Inhibitory and Interfering Substances

Blood samples with high triglyceride levels (>30 g/l) may lead to reduced gDNA yield.

Note: Testing was performed with typical downstream applications to evaluate the quality of extracted nucleic acids. However, different downstream applications may have different purity requirements (e.g., presence of potential inhibitors). Therefore, the identification and testing of relevant substances should also be established as part of the development of any QIAsymphony DSP DNA Midi Kit-related downstream application.

Note: Heparin was not observed to negatively affect performance during the development of the QIAsymphony DSP DNA Midi Kit. However, ISO 20186-2:2019 (E) indicates that heparin in blood collection tubes may affect the purity of isolated nucleic acids and may contaminate eluates, potentially inhibiting certain downstream applications. Therefore, users are responsible for validating whether heparin negatively affects their workflow.

Symbols

This document uses the following symbols. For a complete list of symbols used in instructions for use or on packaging and labeling, refer to the user manual.

Revision History

Revision: R1, June 2022

Description: Version 2, Revision 1

• Updated to Version 2 to comply with IVD
• Added section on Inhibitory and Interfering Substances
• Added section on Eluate Storage
• Added section on Symbols
• Updated Sample Preparation Material section