DNA extraction from a variety of samples using Magnetic Beads
Extracting nucleic acids is a very powerful process that allows us to gain insight into various biological processes for both basic research and clinical applications. DNA isolation (and RNA isolation) is the first step in many modern genomics techniques and applications that require high-quality starting material free of contaminants.
For laboratory managers, complexity remains at the heart of nucleic acid extraction. You could say there are too many and too few options out there. What is the "right" isolation protocol for your sample or application? In this article, we will focus on magnetic bead DNA extraction, which has become one of the most versatile nucleic acid isolation methods.
DNA extraction from a variety of samples using Magnetic Beads
How to Extract DNA (or RNA)
Genomic DNA extraction is the first step in many molecular biology studies and all recombinant DNA techniques. The protocol involves opening the cells and separating the DNA you need from other nucleic acids and cellular components in the sample while keeping it in good condition for downstream analysis. You can take a variety of approaches, from mild to aggressive. Selection depends on several factors, including target DNA, source organism, type and quality of starting material, and application. They typically all share three common steps: lysis, contaminant removal, and DNA recovery.
Step 1: Lysis
Cell lysis involves chemical, mechanical or enzymatic disruption of cell membranes and denaturation of proteins. The exact method depends on your starting material. Bacteria, mammalian cells, plant cells, and human tissues may all require slightly different approaches.
"Gentle" lysis may involve the use of detergents, such as sodium dodecyl sulfate (SDS), or the use of enzymes to break down cell membranes; aggressive lysis may take the form of homogenization to physically disrupt open cell walls.
Step 2: Remove Contaminants
You can use solution-based and solid-phase methods to separate DNA from unwanted lytic debris and potential contaminants. For example, phenol-chloroform DNA extraction separates water-soluble DNA and denatured proteins into distinct phases. This is cheap, but slow and has the potential to affect phenol residues in downstream applications.
Solid phase extraction binds DNA to the surface of a column or bead. For example, silica resin or silica-coated magnetic beads use chaotropic salts to break hydrogen bonds and bind nucleic acids, thereby flushing contaminants away. Oligonucleotide-coated resins can also add a certain degree of specificity, but column kits can quickly add to the cost.
Step 3: Recover Target Nucleic Acids
Downstream applications require your DNA to be in the appropriate form (solvent and concentration). Often, this is just a matter of ethanol-precipitating the DNA, washing, and resuspending in the appropriate buffer. For solid-phase methods, the pH or salt concentration of the buffer first needs to be adjusted to release the nucleic acid.
Magnetic beads for DNA extraction
Because of their versatility and ease of use, magnetic beads provide an excellent alternative to traditional isolation and cleanup methods. They do not require additional centrifugation of samples that may have been agitated, increasing the likelihood of recovering larger fragments, and can be scaled up for higher binding capacity than columns.
Using magnetic beads is simple, no harmful solvents are required, and the release of DNA or RNA requires only adjustment of the properties of the buffer. This simplicity also makes magnetic beads ideal for automation in high-throughput applications.
Professional manufacturer of Nucleic Acid (DNA & RNA) Extraction and Analysis products Magnetic Beads supplier
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