EchoLUTION Cell Culture DNA Kit (50)

For single-step spin column purification from up to 2 x 10⁶ eukaryotic cells (cell lines or primary cells)

The EchoLUTION cell Culture DNA Kit has been developed to purify genomic and mitochondrial DNA from all common cultured cell lines such as HeLa, COS or HEK and also primary cells from eucarytic origin: DNA from up to 2 x 10⁶ fresh or frozen cells is easily purified using the same protocol. No optimization of parameters like lysis time, temperature or elution volume is necessary.

• High DNA yield from purified cells: nearly 100 % recovery from all kinds of cells such as HEK293, HeLa, NIH3T3

• Convenience and speed: complete in just 30 min, no tedious bind-wash-elute procedure

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

Kit contents (50 purifications): 50 EchoLUTION spin columns, Tissue Lysis Buffer, TurboLyse B&C Protease Mix, RNase A, Protector Solution, Clearing solution, Tris Buffer (10x)

Procedure:

Duration:  approximately 30 minutes, including lysis and single step column purification (see flow-chart).

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The initial lysis step takes place under physiological conditions where the enzyme activity of proteases in general is increased. BioEcho’sTurboLyse B&C Protease Mix (patent pending) contains a unique combination of several active enzymes with an expanded substrate specificity. All kinds of cells can be lysed fast and successfully – simple and with the same protocol. The need for mechanical disruption or cell line-specific optimization 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.

All tedious bind-wash-elute steps of common Silica-based procedures are omitted, enabling a significantly faster purification of nucleic acids – 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 from various cell lines (see table below) is up to 2-fold increased per 1 x 10⁶ cells compared to common Silica-based 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. EchoLUTION-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.