Automated purification of high-quality genomic DNA

The Eppendorf epMotion M5073 with MagSep Tissue gDNA kit is an efficient and effective way of isolating genomic DNA in an automated environment.

The MagSep Tissue gDNA Kit has been specifically developed to be used with the Eppendorf epMotion M5073 automated pipetting system. It provides a flexible, easy-to-use solution for magnetic bead-based automated purification from 1-24 samples of high-quality, ready-to-use genomic DNA from a broad variety of sample sources, such as tissue, mouse tails, cultured cells and bacteria.

In this study different mouse tissues were subjected to automated DNA purification using the MagSepTissue gDNA Kit. Genomic DNA purified in this manner showed yields and qualities that were comparable to manual methods and superior to silica column-based technologies. No cross-contamination was detectable and the genomic DNA was directly compatible with downstream real-time PCR amplification. Typical yields ranged from 13.5 μg (10 mg mouse tail material) to 33 μg (10 mg mouse liver).

With the introduction of the epMotion M5073 automated pipetting system with integrated Thermomixer (TMX) and Magnetic Finger Module, a powerful tool for the automation of magnetic bead-based applications became available. The MagSep Tissue gDNA Kit is a suitable addition that enables the user to easily perform hands-free, walk away automated genomic DNA purification. This combination of instrument and kit delivers ready-to-use, high-quality, high-yield genomic DNA that is directly compatible with downstream applications.

To show the effectiveness of the epMotion and MagSep Tissue gDNA kit, 10 mg of mouse tissue was processed for the isolation of genomic DNA. Tissue samples were processed with 25 μL Proteinase K solution and 200 μL lysis buffer, added manually. The lysis was performed overnight in a Thermomixer comfort at 56°C with shaking. After complete lysis, the DNA-containing supernatant was cleared by centrifugation and 225 μL tissue lysate were subsequently transferred into a fresh 2 mL Eppendorf DNA LoBind Tube for subsequent automated processing by the epMotion M5073. For 24 tissue samples the total processing time was approximately 2 hours excluding overnight lysis. The isolation performance of the individual purification methods was assessed by UV measurements of 3 μL and gel electrophoresis of the final eluates.

Depending on the type of tissue, typical DNA yields from 10 mg sample material were in the range of 13.5 to 33 μg for the automated method. DNA purity was good with A260/280 ratios between 1.67 to 1.89.

In a separate experiment, DNA isolated from 24 mouse tail clippings (10 mg each, combined in a master lysate) using the epMotion M5073 showed yields with an average yield of 17.3 μg and CV of 2.09%, giving evidence for a high consistency of the automated method.

The isolation performance of the automated MagSep Tissue gDNA Kit was compared to two widely used silica column-based kits and the manual method using the same reagents. Magnetic bead-based methods (both automated and manual) yielded up to twice as much DNA with purities comparable to the silica column methods, with higher reproducibility of the automated method.

The Eppendorf epMotion M5073 system in combination with the Eppendorf MagSep Tissue gDNA Kit allows the user to conveniently and efficiently purify genomic DNA from tissue and cell samples with minimum hands-on time and a maximum level of automation.

A broad range of sample material can be processed which reproducibly yields large amounts of high-quality, ready-to-use genomic DNA. The safety and reliability of this feature was proven by the fact that no cross-contamination was detectable by real-time PCR.

Visit www.eppendorf.com/epmotion for the full application details.