Nonenzymatic Primer Extension Experiments in the Presence of Coacervates

This article outlines a method for investigating the formation and function of potentially prebiotic complex coacervates, as primitive compartments, generated through electrostatic interactions between anionic nucleic acids and cationic peptides. The Basic Protocols specifically detail the experimental procedures for conducting nonenzymatic RNA primer extension reactions using 2-aminoimidazole-activated ribonucleoside 5'-O-monophosphates in the presence of these coacervates (e.g., composed of specific "host" DNA/RNA sequences and short polyarginine peptide partners). Designed to be accessible to researchers with limited prior experience in coacervate systems, this step-by-step guide provides a practical framework for probing prebiotic polymerization processes, an area of importance in the study of the origins of life. © 2025 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Screening for suitable host DNA/RNA sequences and cationic peptide sequences Basic Protocol 2: Primer extension in the presence of coacervates.