EchoLUTION Cell Culture DNA Kit (250)
- Order number: 010-006-250
- Product weight: 0.662 kg
For single-step spin column purification from up to 2 x 106 eukaryotic cells (cell lines or primary cells)
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High DNA yield from purified cells: nearly 100 % recovery from all kinds of cells such as HEK293, HeLa, NIH3T3
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Convenience and speed: complete in just 30 min, no tedious bind-wash-elute procedure
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Sustainability: 70 % less plastic than with Silica-based methods, no hazardous liquids
Kit contents (250 purifications): 250 EchoLUTION spin columns, Tissue Lysis Buffer, TurboLyse B&C Protease Mix, RNase A, Protector Solution, Clearing solution, Tris Buffer (10x)
EchoLUTION Cell Culture DNA Kit (250) Product no.: 010-006-250
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.
Procedure:
Duration: approximately 30 minutes, including lysis and single step column purification (see flow-chart).
.
The initial lysis step takes place under physiological conditions where the enzyme activity of proteases in general is increased. BioEcho’s TurboLyse 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.
EchoLUTION Cell Culture DNA Kit (250) Product no.: 010-006-250
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.
Procedure:
Duration: approximately 30 minutes, including lysis and single step column purification (see flow-chart).
.
The initial lysis step takes place under physiological conditions where the enzyme activity of proteases in general is increased. BioEcho’s TurboLyse 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.
- BioEcho spin column loading
- EchoLUTION Cell Culture DNA Kit - Product Profile
- Extraction of genomic and plasmid DNA from mammalian cells
- Protocol
SDS (MSDS) saftey data sheet (material safety data sheet), please request: contact@bioecho.de
John Robert Davis, PhD, PostDoc Apoptosis and Proliferation Control Laboratory, The Francis Crick Institute, London, UK
„I used the EchoLUTION CellCulture DNA Kit for extraction of genomic DNA from mammalian cells in culture and it was quick and I obtained higher yields than with the Qiagen kit we use. I have performed a load of PCRs on the gDNA and all working fine. Great kit and will use again!”
Wieland Fahr, Laboratory of Prof. Dr. Stefan Dübel, Department for Biotechnology, Technische Universität Braunschweig, (Germany)
„We used BioEcho’s EchoLUTION kits Tissue DNA Micro and CellCulture DNA for extraction of genomic DNA from cultured cells and from fresh and stabilized human tissue. Both kits impressed with their simplicity, speed and with stable and reproducible yields. Purified DNA could immediately be used for downstream applications such as PCR. With the CellCulture DNA Kit, we were also able to isolate plasmid DNA from eukaryotic cells for efficient subsequent transformation of E. coli.“
Application note:
Extraction of genomic and plasmid DNA from mammalian cells
John Robert Davis, PhD, PostDoc Apoptosis and Proliferation Control Laboratory, The Francis Crick Institute, London, UK
„I used the EchoLUTION CellCulture DNA Kit for extraction of genomic DNA from mammalian cells in culture and it was quick and I obtained higher yields than with the Qiagen kit we use. I have performed a load of PCRs on the gDNA and all working fine. Great kit and will use again!”
Wieland Fahr, Laboratory of Prof. Dr. Stefan Dübel, Department for Biotechnology, Technische Universität Braunschweig, (Germany)
„We used BioEcho’s EchoLUTION kits Tissue DNA Micro and CellCulture DNA for extraction of genomic DNA from cultured cells and from fresh and stabilized human tissue. Both kits impressed with their simplicity, speed and with stable and reproducible yields. Purified DNA could immediately be used for downstream applications such as PCR. With the CellCulture DNA Kit, we were also able to isolate plasmid DNA from eukaryotic cells for efficient subsequent transformation of E. coli.“
Application note:
Extraction of genomic and plasmid DNA from mammalian cells
