EchoLUTION Blood DNA HiYield Kit (10)

for single-step purification of up to 20 µg genomic DNA from 200 – 1000 µl liquid blood... more

EchoLUTION Blood DNA HiYield Kit (10)   Product no.: 010-011-010

for single-step purification of up to 20 µg genomic DNA from 200 – 1000 µl liquid blood ( fresh, frozen, stabilized: Citrat, EDTA, Heparin )

The EchoLUTION Blood DNA HiYield Kit has been developed to variably purify highly pure genomic DNA from 200 µl - 1000 µl blood (fresh, frozen or stabilized). The particularly easy, fast and straight forward procedure is done in just 20 minutes. For larger input volumes up to 1 ml purchasing of costly DNA extraction Midi formats is not necessary.

Specification table EchoLUTION Blood DNA HiYield Kit 

Procedure

Duration:  Done in 20 minutes, including erythrocyte removal, leucocyte lysis and single-step column purification. (see flow-chart).

The EchoLUTION Blood DNA HiYield technology – unlike common bind-wash-elute methods – does not include any inhibitory reagents and avoids harsh high-salt conditions and the use of ethanol. The first step of the procedure comprises a gentle but efficient erythrocyte lysis and hemoglobin removal starting from stabilized or frozen blood. The leucocytes remain intact completely and are subsequently lysed with TurboLyse^TM H Protease solution (patent pending), a unique active enzyme with an expanded substrate specificity. Lysis takes place under physiological conditions where the enzyme activity of proteases is generally increased. Blood samples are lysed fast and successfully – simply and with the same protocol.

At last, DNA is separated from the remaining cellular constituents and from process components via a single centrifugation step. The 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 a 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 EchoLUTION Blood DNA HiYield Kit, extracts up to 20 μg highly concentrated DNA (200 ng/μl) while offering maximal flexibility with 200 μl to 1 ml blood as sample input. For processing larger sample volumes than 200 µl, scientists so far had to rely on costly and inconvenient larger-scale kits (e. g., Midi formats). The standard bind-wash-elute procedures are limited to 200 µl sample volume and suffer from a limited binding capacity, lower DNA concentration and purity (see figure and table).

 

Flexible blood input, high DNA yield, concentration and purity. The indicated volumes of blood (the same donor, 3 samples each) were purified using a common DNA purification kit (Q) or the BE Blood DNA HY Kit (BE). 2 µl eluate were loaded onto an agarose gel; key parameters were measured spectrophotometrically (table); M, DNA size marker, the  largest band representing a 20 kb fragment.

 

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).

EchoLUTION purified genomic DNA is eluted in Tris buffer, pH 7.8 and can immediately be used for downstream applications. 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, 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. NGS detection often requires high DNA concentration and amount. Particularly blood donors with infections, auto-immune diseases or receiving cancer treatment suffer from low leucocyte titer and therefore low amount of DNA. With theEchoLUTION Blood DNA HiYield Kit, these samples result in enriched DNA yield and concentration, enabling sequencing results of the desired coverage and phred score. BioEcho-purified DNA comprises of long DNA fragments, free of contaminants and enzyme inhibitors and is highly suitable for all downstream applications such as PCR, Genotyping and NGS.

 

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