Basics of nucleic acid isolation
Nucleic Acid Isolation is the process by which DNA and/or RNA is separated from proteins, membranes and other cellular material. Nucleic Acids are the key component for many of today’s molecular testing methods, therefore the quality of the isolation step in your process is very important. Since Nucleic Acid Isolation is the first step in the entire molecular process, it is crucial to assure that the yield and purity meet your expectations. These expectations can vary between fields of research and the downstream applications.
There are four basic steps in a Nucleic Acid Isolation procedure, lyse, bind, wash and elute. First, the disruption of the cellular structure to create a lysate. The cell and nucleus are broken open to release the nucleic acid. Secondly, the nucleic acids are bound to a carrier. After the binding step, it is possible to separate the nucleic acid, because after the first lysate step, the nucleic acid has been freed from the nucleus, but it is still mixed with cell debris and other insoluble material. Thirdly, the purification of the nucleic acid of interest to remove any remaining unwanted material, by washing the mixture with specific buffers. At this point the purified nucleic acid is usually re-dissolved in an elution buffer for use in the downstream application and storage.
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