BioEchos genomic DNA extraction technology – New, intelligent and innovative!
Current preparation technology for genomic DNA
Silica-based technologies with their lengthy bind-wash-elute procedure, have been around for many years for all kinds of nucleic acid purification, e. g., for DNA extraction from tissue. These are commonly applied, mainly being a matter of habit, but also because of missing alternatives. However, users experience compromised results in many cases, e.g. with lowered sensitivity and even drop-outs in downstream applications such as PCR. There are two main reasons why results as well as DNA quality of today´s common genomic DNA preparations can be poor: Incomplete lysis of cell walls and harsh, non-physiological purification conditions with the risk of carry-over of organic solvents or salts.
Disadvantages of chaotropic, high-salt working conditions
Fibrous and other hard-to-lyse tissues need to be processed in lengthy protocols and yet result in incomplete lysis, since they are solely based on Proteinase K as the only enzymatic activity for lysis of proteins, larger complexes and cell walls under the applied chaotropic high-salt conditions such as 8 M GuHCl. Also, other sample types such as brain, fibrous and fatty tissues suffer from these inappropriate lysis conditions. The subsequent harsh Silica purification technology binds the DNA onto a membrane in the presence of high concentrations of chaotropic salts and organic solvents and the DNA is eluted with water or aqueous buffers later in the process. This overall procedure introduces shearing forces, interacts with the nucleic acid and - in addition- each bind, wash or elute step is connected to inherent losses in yield. Overall yield and DNA length are lowered, while the risk of co-purifying salts and organic solvents is increased. A compromise in sensitivity for enzyme-dependent analyses such as PCR due to residual inhibiting molecules or just low quantities of DNA is frequently observed. Nevertheless, most labs use this technology, often because of missing alternatives.
BioEcho’s EchoLUTION DNA extraction technology
The BioEcho experts strived to overcome all the described disadvantages with a particularly innovative and convenient technology, that not only significantly improves purity, DNA integrity and yield, but also shortens the overall time to result. The EchoLUTION sample process completely works under aqueous, physiological DNA-protecting conditions, comprising an exceptionally efficient and gentle proprietary lysis followed by a unique single-step purification method based on an optimized principle of reverse chromatography: DNA passes the column matrix without interaction, while impurities are held back and completely removed.
Unmet lysis efficiency
Since the initial lysis step takes place under physiological conditions, the enzyme activity of proteases in general is increased, resulting in a high extraction efficiency in a fraction of time. The TurboLyseTM Protease Mix contains a unique combination of several active enzymes with an expanded substrate specificity and a gDNA protecting component. This proprietary combination drives the overall process: all kinds of cells and tissues can be lysed fast and successfully – effectively and simply with the same protocol. The need for mechanical disruption or lengthy, sometimes over-night lysis has been eliminated.
Genomic DNA purification in just a single-step
The EchoLUTION single-step spin purification procedure combines an initial filtration, holding back potential cell debris, with an extremely efficient reverse purification (spin column, see photo). The cleared lysate is directly loaded onto the spin column. After just one single spin, the purified high molecular weight genomic DNA is eluted, while all debris and cellular compartments, inhibitors, small fragments (< 50 bp), nucleotides, and salts are completely removed. Impurities are held back by the purification matrix and the highly pure DNA is collected in the flow-through. The purified DNA is easily recovered with close to 100 % efficiency and does not contain residual traces of substances which interfere with enzyme-dependent downstream applications or falsify OD measurements.
Convenient workflow and higher DNA quality
The BioEcho procedures further stand out in being short and convenient: the EchoLUTION protocols are up to 3 times faster than common Silica-based methods. Centrifugation steps are reduced from 8 to a maximum of 3, overall handling steps from >20 to 8, protocol duration from several hours to 30 minutes. One protocol can be applied for all kinds of tissues or cells, making lab life much easier than before. Plus, the risk of chemical and physical DNA damage is strongly reduced, e. g. by shearing of high molecular weight molecules – target nucleic acids remain untouched throughout the entire workflow.
High performance and reliable results with BioEcho DNA extraction Kits
Since EchoLUTION purified DNA is completely free of contaminants and enzyme inhibitors like chaotropic reagents and organic solvents and furthermore composed of particularly intact DNA fragments, it is highly suited for all downstream applications. Particularly in experiments, where sensitivity is of high importance such as qPCR with limited sample material, EchoLUTION purified DNA outperforms results obtained with silica technology purified DNA. (see qPCR figure)
Responsible & sustainable
Since GuHCl is irritant, aggressive and corrosive, it must be used with caution, and contaminated plastic waste and liquids must be disposed of separately – at high costs! It is part of BioEcho’s vision to develop sustainable, eco-friendly lab processes with reduced amount of plastic-based kit components that are consumed during the lab process. Due to the convenient single-step purification technology, 70% less plastics are wasted than with its Silica counterparts, and no hazardous liquids are required. BioEcho´s Kit and spin column packaging is completely free of plastics, just based on carton or corn starch, thus additionally conserving natural resources. The decision to use BioEcho products will be beneficiary for lab safety and health as well as for financial resources.
BioEchos EchoLUTION technology for purification of genomic DNA combines a particular effective lysis with an outstandingly convenient and fast spin column purification. The gentle yet powerful workflow results in highly pure and intact genomic DNA. It is completely free of contaminants and enzyme inhibitors like chaotropic reagents and organic solvents and is highly suited for all downstream applications such as PCR, qPCR, Genotyping and NGS applications.