The invention discloses a method for complete cryopreservation and thawing of an RNC (intracellular ribosome nascent-chain complex). The method comprises the following steps: adding a translation elongation inhibitor into cell culture fluid in normal growth, cooling cells to the temperature of 0-4 DEG C after the translation elongation inhibitor effectively acts on the RNC and translation elongation is inhibited, cleaning the cells by using a PBS solution which is precooled to the temperature of 0-4 DEG C, adding the PBS solution into the cells, rapidly freezing in liquid nitrogen, preserving at an ultra-low temperature, thawing on ice, and extracting the RNC. According to the method, ribosome is effectively fixed by adopting the translation elongation inhibitor and is unlikely to dissociate, and the translation conditions in the cells are fixed. Moreover, the RNC is effectively cryopreserved for long-term storage and transport through a liquid nitrogen quick freezing method, and the extracted RNC after thawing is the same as the RNC extracted from fresh cells, so the limitation of time and space is broken through, and convenience is provided for research of the RNC and development of commercial service of the RNC.