Step-by-step guide for effective genomic DNA purification using SPINeasy DNA extraction kit:
Genomic DNA purification is a vital step in various scientific applications. Purifying genomic DNA essentially means isolating it from other cellular components and contaminants for downstream applications. There are many methods of genomic DNA purification, such as Phenol-Chloroform Extraction, Magnetic Bead-Based Technique and Spin Column-Based Kits.
In this blog, we will explore the most effective method of genomic DNA purification: the SPINeasy® DNA extraction kits.
The SPINeasy® DNA purification kit is designed specifically for rapid and efficient genomic DNA purification from various samples. Here is a general protocol for purifying DNA using MP Biomedicals’ SPINeasy® DNA purification kits and instruments.
Cell lysis: Cell Lysis is the process of splitting the cell open. A chemical or mechanical method disrupts the membrane of the cell and releases the cellular contents, including the genomic DNA, in a hypotonic solution (e.g., diluted sucrose solution).
During chemical cell lysis, some common chemicals including organic solvents (alcohols, ethers), chelating agents, detergents and chaotropic agents (urea, guanidine) are used.
Transfer to Spin Column: The lysate which is mixed with the binding solution, is then transferred to a spin column. Spin columns are small tubes containing a silica-gel membrane that selectively binds to nucleic acids. The binding solution contains a mixture of buffer and ethanol/isopropanol. This solution is designed to create conditions where nucleic acids bind to a silica-gel membrane. The buffer maintains the optimal pH and salt concentration for nucleic acid to bind onto the silica membrane.
Centrifugation: The spin column is placed in a centrifuge. When the centrifuge is activated, it generates a centrifugal force that drives the binding solution through the silica-gel membrane. As the solution passes through the membrane, nucleic acids bind to the silica while other contaminants are washed away.
Washing: Additional washing steps are often performed to remove impurities after the initial centrifugation step. These wash steps involve the addition of specific wash buffers onto the spin column, followed by centrifugation. The wash buffers help to remove salts, proteins, and other contaminants, leaving only nucleic acids bound to the silica membrane.
Elution: Finally, purified nucleic acids are eluted from the silica membrane using low-salt buffer or water. Elution disrupts the binding between nucleic acids and the silica, allowing the purified nucleic acids to be collected in a separate tube.
At MP Biomedicals, M.P. Biomedical provides different kinds of SPINeasy® DNA purification kits to suit any of your sample types. Explore the wide range of Spin Column-based Extraction Kits by MP Biomedicals.
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