A buffer solution formulated for the efficient extraction of RNA from cells or tissues under aqueous conditions generally contains a mixture of chemical compounds designed to disrupt cellular structures while preserving the integrity of the RNA molecules. Common components include detergents, such as sodium dodecyl sulfate (SDS) or Triton X-100, which solubilize cell membranes and denature proteins. Chaotropic agents, like guanidinium thiocyanate or urea, are often incorporated to further denature proteins and inhibit RNases, enzymes that degrade RNA. Additionally, the solution typically contains a buffering agent, such as Tris-HCl, to maintain a stable pH, which is crucial for RNA stability. Ethylenediaminetetraacetic acid (EDTA) may also be present to chelate divalent cations, inhibiting DNases and RNases that require these ions for activity. Salt, such as sodium chloride, may be included to optimize the binding of RNA to silica-based purification columns if used in downstream processing.
The use of such a solution is paramount in molecular biology workflows where high-quality RNA is essential for downstream applications. Obtaining intact and pure RNA is critical for accurate and reliable results in techniques like reverse transcription PCR (RT-PCR), RNA sequencing, and microarray analysis. Prior to the development of effective lysis buffers, the isolation of RNA was a laborious and often unreliable process, prone to degradation. The advent of optimized aqueous solutions for cell lysis has greatly improved the efficiency and reproducibility of RNA extraction, enabling significant advances in gene expression studies and other related research areas.
