EchoLUTION Plant DNA Kit (50)

For single-step spin column purification of genomic DNA from up to 50 mg of plant tissue (leaves, soft roots, seeds, fruit tissue, bud & blossom tissue)

• High DNA yield: up to 5-fold more than Silica plant kits
• High purity: reliable removal of inhibitors for downstream reactions (e. g., PCR, NGS)
• Convenience: no tedious bind-wash-elute procedure, grinding solution enables faster disruption of samples
• Speed: Whole process complete in 45 min
• Sustainability: 70 % less plastic than with Silica-based methods, no hazardous liquids

Kit contents (50 purifications): 50 EchoLUTION spin columns, Plant Lysis Buffer, TurboLyse P Protease, RNase A, Grinding Suspension, Clearing Solution, Tris Buffer (10x)

The EchoLUTION Plant DNA Kit has been developed to quickly purify genomic DNA from various species and types of tissues, including leaf, seed, fruit and root material (see table). No optimization of parameters like lysis time, temperature or elution volume is necessary.

Procedure:

Duration:  Done in 45 minutes, including tissue lysis and single-step column purification. (see flow-chart).

Grinding solution (provided) enables a fast and convenient disruption of plant tissue when not using bead beating. The subsequent lysis step takes place under physiological conditions where the enzyme activity of proteases is generally increased. BioEcho’s TurboLyse P Protease Mix (patent pending) contains a unique combination of several active enzymes with an expanded substrate specificity. All kinds of plant tissues can be lysed fast and successfully – simply and with the same protocol. BioEcho also provides tissue grinding pestles supporting the initial mechanical disruption step in a 1.5 ml reaction tube.
The subsequent EchoLUTION single-step spin purification procedure combines an initial filtration, holding back potential cell debris, with an extremely efficient reverse purification. The cleared lysate is directly loaded onto the spin column. After just one single spin, the purified high molecular weight genomic and mitochondrial DNA is eluted, while all debris and cellular compartments, inhibitors, small fragments (< 50 bp), and salts are completely removed. Since impurities are held back by the purification matrix and since the highly pure DNA is collected in the flow-through, the complete DNA is easily recovered with close to 100 % recovery and highest purity. Addition of carrier nucleic acids is not required since matrix interactions have been minimized.

The extraction efficiency of the EchoLUTION Plant DNA Kit process is up to 5 fold higher than with common Silica-based purification kits (see below).

All tedious bind-wash-elute steps of common Silica-based procedures are omitted, enabling a significantly faster purification of nucleic acid samples, with less handling, drastically reduced plastic consumable usage and no use of hazardous materials like chaotropic salts or organic solvents (see fig).

Plastic waste produced from a 50 reactions Silica-based DNA kit and from an EchoLUTION kit (including kit components and consumables that are not part of the kit). Bags contained in BioEcho kits are cellophane-based. In numbers:
 

Purified genomic DNA is eluted in Tris buffer, pH 7.8 and can immediately be used for downstream applications (see below). The amount obtained (see table) is up to 10-fold increased per mg tissue compared to common Silica methods. EchoLUTION purified genomic DNA preparations usually show an A260/280 ratio of 1.8 to 2.0.
 

Applications

Since EchoLUTION kit purified genomic DNA does not contain any traces of chaotropic salts, organic solvents, heavy metal ions or other inhibitory impurities, a superior performance is seen in downstream applications such as real-time PCR or PCR (see figure), even with undiluted samples with the DNA solution making up to 45 % of the total PCR reaction volume. This advantage can make a huge difference, e. g., for small tissue samples with corresponding low quantity of purified DNA or when highest sensitivity is required. BioEcho-purified DNA comprises of particularly long DNA fragments, free of contaminants and enzyme inhibitors like chaotropic reagents and organic solvents and is highly suitable for all downstream applications such as PCR, Genotyping and NGS.