RNA extraction isolates RNA strands for genetic research, allowing scientists to study the pathway from DNA to protein production and cellular function. Different tissue and cell types require different RNA extraction methods, and RNA degradation is a common complication that can render a sample useless.
Ribonucleic acid (RNA) extraction is a commonly used laboratory process in molecular biological research. By isolating RNA strands, researchers can study the genetic components of a variety of organisms, from bacteria to humans. Although deoxyribonucleic acid (DNA) is the most common nucleic acid associated with genetic research, RNA is the connection between DNA and cellular proteins responsible for a variety of structural roles. RNA extraction allows scientists to study the pathway from DNA information to protein production and cellular function.
Nucleic acids, in the form of DNA and RNA, contain the genetic information necessary for cell function and replication. Ribonucleic acid, or RNA, is the link between DNA and cellular proteins. Specific types of RNA copy specific sections of DNA to help make these proteins. Tracing the pathway between DNA and cellular proteins helps researchers understand gene regulation and protein synthesis within individual cells. Processes that facilitate RNA extraction and ensure sample stability provide researchers with a means to follow the pathway between DNA and RNA.
During RNA extraction, biological samples are reduced and purified to such an extent that the RNA is isolated for later examination. Molecular components, such as proteins, RNA, and DNA, break down or isolate at different times, allowing specific types of RNA to be extracted. Different tissue and cell types require different RNA extraction methods. While there are a variety of methods for performing RNA extraction, the most common uses centrifugation to mix biological samples with various chemical compounds, collectively known as denaturants.
A common method of RNA extraction is phenol-chloroform extraction, also known as PC or PCIA. Using this method, tissue samples are mixed with equal parts of phenol and chloroform. Known as a two-phase blend, the solution is centrifuged in a denaturing solution. Two phases result, the first being the aqueous phase and the second being the organic phase. It is during the aqueous phase that the RNA extraction takes place, with the help of ethanol precipitation.
Other methods for RNA extraction vary depending on sample size, tissue type, and the need for full or partial RNA. For yeast, plant or animal samples, complete RNA or total eukaryotic RNA, samples are required. Bacteria, on the other hand, require total prokaryotic RNA. Each type of RNA requires a different method of sample preparation, extraction, and storage.
Complications with RNA extraction are common and usually involve RNA degradation after extraction. Ribonuclease enzymes commonly found in tissue samples rapidly degrade RNA. If not used quickly, RNA degradation can render a sample useless. In response, many RNA extraction methods include steps for preparing samples for storage, both before and after extraction, to reduce or slow RNA breakdown.
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