Application Note

AUTOMATED DNA ISOLATION FROM ZYMO RESEARCH

ZR-96 Genomic DNA ™ MagPrep Catalog Nos. D3083 & D3084 High-throughput, automated DNA recovery from whole blood, biological fluids, cells and tissue with the Tecan Freedom EVO ®.

Introduction The ZR-96 Genomic DNA ™ MagPrep features a simple, high throughput (96-well) procedure for the rapid isolation of total DNA (e.g., genomic, mitochondrial, viral) from a variety of biological sample sources. Compatible sample sources include whole blood, bodily fluids, cell suspensions, tail snips, ear punches, muscle biopsies, etc. This product has been optimized for maximal recovery of ultra-pure DNA without RNA contamination. The extraction procedure includes Proteinase K for thorough sample lysis and features high efficiency MagBinding Beads for DNA purification. PCR inhibitors are effectively removed during the purification process. The purified DNA is suitable for PCR, nucleotide blotting, DNA sequencing, restriction endonuclease digestion, bisulfite conversion/methylation analysis, and other downstream applications.

The platform and procedure are easy to set up to obtain consistent, reliable DNA recovery using the Tecan Freedom EVO®. Automating DNA purification allows researchers to obtain reliable user-independent results, increasing throughput and consistency.

Application Note

AUTOMATED DNA ISOLATION FROM ZYMO RESEARCH

Automation Equipment

Overview of Procedure

Samples are digested with a highly active Proteinase K, then Genomic Lysis Buffer and MagBinding Beads are dispensed into the sample for DNA binding. The bound DNA is washed several times and eluted with the provided DNA Elution Buffer or water. The simplicity of the procedure facilitates rapid turnaround times for the purification of high-quality genomic DNA.

• Tecan Freedom EVO® • Freedom EVOware® • 8 channel Liquid Handling Arm (LiHa),

configured for Disposable Tips (DiTis) • Robotic Manipulation Arm (RoMa) • Te-Shake™ Shaker • 96-well Magnetic Stand

Figure 1. Example deck layout

Bind and wash DNA. Remove lysate and wash. Elute purified DNA.

Application Note

AUTOMATED DNA ISOLATION FROM ZYMO RESEARCH

Results

Figure 2. Comparison between manual and automated processing . 50 µl porcine blood was sampled and magnetic bead purification performed manually or using the Tecan Freedom EVO® 150. While there is ~30% less yield, the comparison demonstrates the automated liquid handling platform to produce more consistent yields and recovered volumes.

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B

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Automation Manual

Concentration (ng/µl) Volume (µl) Yield (ng x10)

Figure 3. High molecular weight DNA is ideal for downstream applications including PCR. Panel A. Both automated and manual methods produce the same expected recovery from 50 µl porcine blood. Panel B. A portion of the eluate was used for PCR, generating the expected product from the beta-actin gene. The DNA is resolved in a 2% TAE/EtBr agarose gel

Application Note

AUTOMATED DNA ISOLATION FROM ZYMO RESEARCH

Conclusions High quality DNA (A260/A230 > 1.8, A260/A280 > 1.8) is recovered with consistent yields using the ZR-96 Genomic DNA™ MagPrep and Tecan Freedom EVO® platform. Isolated genomic DNA is an ideal substrate for downstream applications such as PCR, sequencing, restriction endonuclease digestion, bisulfite conversion/methylation analysis, etc. This streamlined procedure is simple, hassle-free, and only requires standard automation equipment. Specifications

• Sample Sources – Whole blood, plasma, or serum from humans, mice, rats, etc. Also, tissue, cultured cells, as well as a variety of biological liquids are effectively processed using this kit. Solid tissues such as tail snips, ear punches, muscle biopsies, etc. are also compatible.

• DNA Size Limits – Capable of recovering genomic DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered.

• DNA Recovery – Up to 10 µg/well total DNA is eluted into ≥ 50 µl DNA Elution Buffer or water.

• Product Detergent Tolerance – ≤ 5% Triton X-100, ≤ 5% Tween-20, ≤ 5% Sarkosyl, ≤ 0.1% SDS.

Products

Product Description Catalog No. Kit Size (Preps.)

ZR-96 Genomic DNA ™ MagPrep D3083 D3084

2 x 96 4 x 96