The invention discloses a preparation method of a cold shock protein. The preparation method is characterized by comprising the following steps of 1, TRIzol method-based total RNAs extraction: grinding a mouse tissue sample by a mortar and adding 1ml of a pyrolysis liquid into each 50-100mg of the tissue, 2, cDNA first-chain synthesis: adding 5 microliters of a RNA solution, 1 microliter of Oligo and 1 microliter of dNTP into 20 microliters of a reaction system and carrying out a reaction process at a temperature of 65 DEG C for 5min, 3, a reaction process: adding 1 microliter of cDNA, 2 microliters of dNTP, 1 microliter of TapDNA polymerase, 1 microliter of an upstream primer, 1 microliter of an downstream primer and 2 microliters of 10*buffer into 20 microliters of the reaction system, 5, inducible expression, 6, preliminary purification and 7, high-degree purification. The cold shock protein which is coldinducible RNA-binding protein (CIRP) has effects of protecting nervous tissue cells and resisting nervous tissue cell apoptosis.