The article provides a comprehensive protocol for the extraction and processing of silk fibroin from Bombyx mori cocoons to fabricate various biomaterials. Silk fibroin, derived from the cocoon of the silkworm, is a widely studied protein polymer with excellent mechanical properties, biocompatibility, and controllable degradation rates. The protocol includes methods for extracting silk fibroin, concentrating it, and processing it into different materials such as hydrogels, tubes, sponges, composites, fibers, microspheres, and thin films. These materials can be used for implants, tissue engineering, disease models, and drug delivery. The extraction process involves boiling the cocoons to remove sericin, followed by dissolution in lithium bromide solution and dialysis to obtain pure silk fibroin. The solution can be lyophilized or concentrated for storage and further processing. The article also details specific protocols for creating various silk-based materials, including dipping and gel spinning for tubes, vortexing and sonication for gels, electrospinning for fibers, and casting for films and microspheres. Each step is accompanied by critical and troubleshooting tips to ensure successful material fabrication.The article provides a comprehensive protocol for the extraction and processing of silk fibroin from Bombyx mori cocoons to fabricate various biomaterials. Silk fibroin, derived from the cocoon of the silkworm, is a widely studied protein polymer with excellent mechanical properties, biocompatibility, and controllable degradation rates. The protocol includes methods for extracting silk fibroin, concentrating it, and processing it into different materials such as hydrogels, tubes, sponges, composites, fibers, microspheres, and thin films. These materials can be used for implants, tissue engineering, disease models, and drug delivery. The extraction process involves boiling the cocoons to remove sericin, followed by dissolution in lithium bromide solution and dialysis to obtain pure silk fibroin. The solution can be lyophilized or concentrated for storage and further processing. The article also details specific protocols for creating various silk-based materials, including dipping and gel spinning for tubes, vortexing and sonication for gels, electrospinning for fibers, and casting for films and microspheres. Each step is accompanied by critical and troubleshooting tips to ensure successful material fabrication.