[en] Grey oyster mushroom (Pleurotus pulmonarius), black jelly mushroom (Auricularia auricula) and paddy straw mushroom (Volvariella volvacea) are the most common and typical edible mushrooms species enjoyed by Malaysians owing to their good flavors properties and high nutrients content. Master culture or commonly known as mother culture is a crucial element in producing a good mushroom seedlings. In vitro technique of initiating tissues in preparing the mother culture is the most crucial step. Firstly, screening and selection for quality fruiting body used to initiate tissue must be fresh, healthy and free from any spots of pest and diseases. Secondly, place in doing the activity must be closed environment, clean and comfortable such as in laminar air flow or clean chamber. Preparation for the clean area is conducted 30 minutes prior to tissue excises. Thirdly, tools in dissecting the tissues must be dry, sterile and normally autoclaved at 121 degree Celsius, 15 psi for 15 minutes. Besides, media used in growing the mycelium should be freshly prepared and free from any spot from bacteria and/or fungus. Fourthly, personnel carrying out the activities must be clean, equipped with laboratory coat and healthy. Fifthly, the process in excising the tissues should be conducted carefully and gently. All steps mentioned are important in getting a good material free from contamination. This paper discusses in depth the steps and procedures taken in initiating different types of mushroom. (author)