This protocol introduces soft lithography, a collection of techniques based on printing, molding, and embossing with an elastomeric stamp. Soft lithography is versatile, cost-effective, and suitable for a wide range of materials, including biological applications. It is particularly useful for creating three-dimensional and curved structures, generating well-defined surface chemistries, and is compatible with various applications such as cell biology, microfluidics, lab-on-a-chip systems, and flexible electronics/photronics. The protocol focuses on three commonly used soft lithographic techniques: microcontact printing (μCP) of alkanethiols and proteins on gold-coated and glass substrates, replica molding for fabricating microfluidic devices in poly(dimethyl siloxane) (PDMS), and solvent-assisted micromolding of nanostructures in poly(methyl methacrylate) (PMMA). The procedures for designing patterns, fabricating masks and masters, and creating elastomeric stamps are detailed, along with specific steps for μCP, replica molding, and solvent-assisted micromolding. The protocol also includes troubleshooting tips and anticipated results, emphasizing the advantages of soft lithography over traditional photolithography in terms of cost, flexibility, and material compatibility.This protocol introduces soft lithography, a collection of techniques based on printing, molding, and embossing with an elastomeric stamp. Soft lithography is versatile, cost-effective, and suitable for a wide range of materials, including biological applications. It is particularly useful for creating three-dimensional and curved structures, generating well-defined surface chemistries, and is compatible with various applications such as cell biology, microfluidics, lab-on-a-chip systems, and flexible electronics/photronics. The protocol focuses on three commonly used soft lithographic techniques: microcontact printing (μCP) of alkanethiols and proteins on gold-coated and glass substrates, replica molding for fabricating microfluidic devices in poly(dimethyl siloxane) (PDMS), and solvent-assisted micromolding of nanostructures in poly(methyl methacrylate) (PMMA). The procedures for designing patterns, fabricating masks and masters, and creating elastomeric stamps are detailed, along with specific steps for μCP, replica molding, and solvent-assisted micromolding. The protocol also includes troubleshooting tips and anticipated results, emphasizing the advantages of soft lithography over traditional photolithography in terms of cost, flexibility, and material compatibility.