EchoLUTION Tissue DNA Micro Kit (10)

For single-step spin column purification of genomic DNA from 0.1 to 10 mg human or animal tissue (fresh, frozen, or stabilized)

• High DNA yield from small samples: nearly 100 % recovery, no loss of precious sample

• Convenience and speed: complete in less than 40 min, no tedious bind-wash-elute procedure

• Sustainability: 70 % less plastic than with Silica-based methods, no hazardous liquids

Kit contents (10 purifications): 10 EchoLUTION spin columns, Tissue Lysis Buffer, TurboLyse T Protease Mix, RNase A, Protector Solution, Clearing solution, Tris Buffer (10x)

The EchoLUTION Tissue DNA Micro Kit has been developed to purify genomic and mitochondrial DNA from all kind of tissues, it has been particularly developed for small input samples. 0.1 to 10 mg fresh, frozen or stabilized tissue and also fresh and frozen DNase-rich tissues – such as liver, kidney and intestine – are easily purified using the same protocol. No optimization of parameters like lysis time, temperature or elution volume is necessary.

Procedure:

Duration:  less than 40 minutes, including tissue lysis and single-step column purification. (see flow-chart).

The initial lysis step takes place under physiological conditions where the enzyme activity of proteases is generally increased. BioEcho’s TurboLyse T Protease Mix (patent pending) contains a unique combination of several active enzymes with an expanded substrate specificity. All kinds of tissues can be lysed fast and successfully – simply and with the same protocol. The need for mechanical disruption or lengthy, sometimes over-night lysis has been eliminated.

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 (< 300 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. This overall procedure is highly suitable for small samples like biopsies or tissue from small animals as those are particularly precious and DNA losses are reduced to a minimum. Addition of carrier nucleic acids is not required since matrix interactions have been minimized.

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 5-fold increased per mg tissue compared to common Silica methods. Protector Solution is supplied to enable long-term storage of DNA samples. EchoLUTION purified genomic DNA preparations usually show an A260/280 ratio of 1.8 to 2.0.
 

Applications

Since EchoLUTION kit purified genomic DNA do 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, 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 as in certain diagnostics applications. 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 applications.