The invention provides an incubation material used for turtles and soft-shelled turtles. The incubation material used for the turtles and the soft-shelled turtles mainly comprises wood chips, dahurian patrinia herb, humifuse euphorbia herb, honeysuckle, paris polyphylla, spreading heydyotis herb, poria cocos, barbed skullcap herb, lobelia chinensis, corydalis bungeana, sophora flavescens, dandelion, gallnut, pericarpium granati, hawthorns and plantain herb. The invention further provides a preparation method of the incubation material used for the turtles and the soft-shelled turtles. The preparation method comprises the steps of cutting all medicinal materials into pieces, mixing the medicinal materials to be uniform, then heating and decocting the medicinal materials into water apozem, then utilizing ethyl alcohol for conducting the sedimentation treatment, after the apozem is concentrated or dried, conducting the separation and purification, adding the apozem into the wood chips for standing for 3-4 days, then laying the apozem under the blazing sun for a long time, or conducting the drying, and finally after the sterilization is conducted, obtaining a finished product of the incubation material used for the turtles and the soft-shelled turtles. The incubation material used for the turtles and the soft-shelled turtles is free of drug resistance, residual, side effect and environmental pollution, obvious in anti-bacteria and sterilization effect, low in cost and capable of effectively preventing and curing diseases which the turtles and the soft-shelled turtles often suffer from such as the necrotic-skin disease, the red-neck disease and the parotitis.