Extracting high-quality genomic DNA from whole blood is the cornerstone of successful molecular diagnostics. Whether you're conducting PCR, sequencing, or downstream genomic analyses, the integrity and purity of DNA directly impact your results. Automation systems like the Manta offer a robust solution, streamlining the extraction process with high reproducibility, improved throughput, and minimized human error. The Manta automated extraction system is engineered specifically to tackle these challenges head-on. By seamlessly integrating enzymatic digestion, magnetic bead-based purification, and precise fluid handling, Manta delivers reproducible, high-purity genomic DNA with minimal hands-on intervention. The system is designed to standardize extraction across a diverse range of blood sample conditions — from freshly drawn EDTA tubes to frozen or even hemolyzed specimens — ensuring that each run meets rigorous quality standards required for clinical decision-making.

In this article, we dive deep into the exact, step-by-step protocol for extracting genomic DNA from whole blood using Manta.

Overview of Kit Components

At the heart of the Manta extraction workflow is a kit containing all necessary reagents and consumables, designed for a seamless end-to-end procedure. A key reagent in this process is Proteinase K, an endolytic serine protease enzyme that digests proteins and nucleases in the blood lysate, thereby protecting the DNA from degradation. It is supplied in a lyophilized form for stability and reconstituted before use. The reconstitution involves adding a precise volume of Proteinase K Diluent, after which the enzyme solution is stored at -20°C to maintain activity. This stability over multiple freeze-thaw cycles ensures consistent enzymatic activity batch after batch, crucial for the reliable digestion of blood proteins.

The Blood Lysis Buffer (BL) is a carefully formulated detergent-based solution that disrupts red and white blood cell membranes, facilitating the release of genomic DNA. Proper lysis is essential to maximize DNA yield and prevent contamination by cellular debris.

The Manta system uses a specially designed cartridge for each sample, which contains six wells pre-loaded with reagents. These wells have distinct roles: a binding buffer facilitates the adsorption of DNA onto magnetic beads; three sequential wash buffers remove proteins, salts, and other impurities; and an elution buffer releases the purified DNA from the beads into the final collection well. This compartmentalized approach allows the system to automate complex fluidic handling with precision and repeatability.

Pre-Digestion Step: Optimizing Protein Degradation and Lysis

The pre-digestion phase is vital to break down protein contaminants and facilitate cell lysis. In this step, 30 μL of reconstituted Proteinase K is combined with 200 μL of whole blood and an equal volume of Blood Lysis Buffer in a microcentrifuge tube. Thorough mixing by vortexing ensures homogeneous distribution of enzymes and lysis buffer, promoting efficient digestion of histones and nucleases bound to genomic DNA.

Incubation at 70°C for 10 to 12 minutes enhances enzymatic activity and accelerates protein digestion. This elevated temperature also helps denature proteins and facilitates cell membrane disruption. The precise control of temperature during incubation is critical; too low, and enzymatic activity diminishes, risking incomplete digestion; too high, and DNA degradation might occur.

For samples that are hemolyzed or inherently challenging, the protocol recommends the addition of an LE Buffer (SKU-CBWC162). This buffer acts to further stabilize nucleic acids and improve the overall yield by enhancing lysis conditions.

Cartridge Preparation: Ensuring Optimal Reagent Delivery

Each cartridge is pre-loaded with carefully measured volumes of binding buffer, magnetic beads, wash buffers, and elution buffer. Before use, gentle vortexing and tapping of the cartridge ensure all reagents are settled at the bottom of their respective wells, preventing cross-contamination or incomplete reagent delivery during the automated run.

The critical step involves transferring the pre-digested lysate to Well 1, which contains the binding buffer. The binding buffer’s composition — often containing chaotropic salts — facilitates the adsorption of DNA onto the magnetic beads in the subsequent step. Proper mixing with a pipette is essential to allow full interaction between the DNA and binding reagents, maximizing capture efficiency.

The cartridges are then loaded into the Manta’s deck tray, ensuring proper orientation and seating. This careful handling is crucial for the magnetic bead-based extraction mechanism to function smoothly.

Running the Automated Extraction

• Select Open Door on the Manta's touchscreen.

• Remove the tray, place it in a biosafety cabinet, and add 430 μL of lysate to Well 1 (binding buffer well).

• Fit magnetic sleeves into the machine with an audible click.

• Place the tray back into the Manta, ensuring proper cartridge placement.

• Choose ‘Choose extraction protocol’ on the screen.

• Select the protocol labeled ‘CB-200-J3’ and hit Continue.

• Once the run completes, collect the eluted DNA from Well 6 into a DNAse-free microcentrifuge tube.

• Store extracted DNA at -20°C for long-term preservation.

Why Choose Manta for Whole Blood DNA Extraction?

• Reduced hands-on time: Less than 10 minutes of active handling, freeing lab personnel for other tasks.

• Fast on-deck runtime: Approximately 28 minutes per extraction cycle.

• Flexible batch sizes: Run even a single sample without waiting for large batches, ideal for clinical labs with variable throughput.

• Optimized reagents and cartridge design: Deliver consistent high yields and purity.

• Reproducible results: Average DNA yield ranges from 80 to 150 ng, with purity ratios (A260/280) between 1.7 and 1.8, and (A260/230) between 2.0 and 2.2, suitable for sensitive downstream applications.

• Scalable protocols: High-volume extraction options are also available for larger throughput needs. Get it here

Tips for Getting the Best Results with Manta

• Always use fresh or properly stored blood samples.

• Avoid repeated freeze-thaw cycles.

• Follow the incubation times and temperatures precisely.

• Handle cartridges carefully to prevent reagent spillage.

• Use DNAse-free tubes and tips to avoid contamination.

• Regularly maintain and sterilize the Manta system.

See how Manta can seamlessly fit into your lab. Watch the Manta in action: Virtual tour of Manta - Nucleic acid isolation machine.