Genes · 2025
The Interference of RNA Preservative and Post-Collection Interval on RNA Integrity from Different Mice Tissues.
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Xie Ting, Zhu Hui, Wang Xiaoxi, Li Fangyuan, Wang Anqi, Zhang Yaran, Zhang Sumei, Guo Dan
Аннотация
BACKGROUND: For precise and reliable gene expression analysis, the acquisition of high-quality RNA is contingent upon excellent tissue preparation and handling. The optimal method for preserving tissues after surgical resection remains challenging due to the delays in delivery or the absence of cold storage equipment. Although RNAlater has been extensively adopted for tissue preservation, few studies have systematically evaluated the effects of various tissue preservation solutions and post-collection intervals on RNA integrity across a range of tissue types.
METHODS: Ten types of mouse tissues, representing common tissue species in biobanks, were collected after resection. Tissues were either flash-frozen in liquid nitrogen as controls or immersed in one of three RNA preservatives-TRIzol and two commercial RNAlater solutions-and stored at room temperature (RT) for 0, 4, or 8 h before being frozen. Total RNA was extracted using TRIzol method, and its integrity was assessed using the RNA Integrity Number (RIN).
RESULTS: The results indicated that both the post-collection interval and the type of RNA preservative significantly impact RNA integrity. Pancreatic tissue showed the poorest RNA integrity (RIN < 5.5), whereas heart and ovary tissue yielded high-quality RNA (RIN > 7) even without any preservatives after 8 h at RT. To maintain baseline RNA integrity (RIN > 5.5), tissues including brain, kidney, muscle, liver, intestine, and uterus should be immersed in preservative and frozen within 8 h. For lung tissue preserved in RNAlater, the maximum recommended time at RT was 4 h.
CONCLUSIONS: Robust, high-quality RNA can be obtained from most mouse tissues stored in RNA preservatives for up to 8 h at RT, with only minor variations observed across the different preservatives tested.
METHODS: Ten types of mouse tissues, representing common tissue species in biobanks, were collected after resection. Tissues were either flash-frozen in liquid nitrogen as controls or immersed in one of three RNA preservatives-TRIzol and two commercial RNAlater solutions-and stored at room temperature (RT) for 0, 4, or 8 h before being frozen. Total RNA was extracted using TRIzol method, and its integrity was assessed using the RNA Integrity Number (RIN).
RESULTS: The results indicated that both the post-collection interval and the type of RNA preservative significantly impact RNA integrity. Pancreatic tissue showed the poorest RNA integrity (RIN < 5.5), whereas heart and ovary tissue yielded high-quality RNA (RIN > 7) even without any preservatives after 8 h at RT. To maintain baseline RNA integrity (RIN > 5.5), tissues including brain, kidney, muscle, liver, intestine, and uterus should be immersed in preservative and frozen within 8 h. For lung tissue preserved in RNAlater, the maximum recommended time at RT was 4 h.
CONCLUSIONS: Robust, high-quality RNA can be obtained from most mouse tissues stored in RNA preservatives for up to 8 h at RT, with only minor variations observed across the different preservatives tested.