| With the rapid growth and development of academic courses,such as precision medicine and translational medicine,which requires human biological resources as the main resourcesan increasing number of biobanks have been built around the world in recent years.Scientists and researchers are paying more and more attention to the long-term green preservation of biological samples in an economical and safe way.At present,the preservation of RNA samples in biobanks at home and abroad mainly depends on ultra-low temperature storage,which is not only expensive,but also poses threat to energy conservation and environmental protection,moreover,the risk of power failure for ultra-low temperature storage system may also exist.Any of the factors mentioned above will result in the degradation of RNA samples.Therefore,a substitute RNA preservation solution which is reliable and environmentally friendly is urgently needed in current biobanks.The room temperature preservation solution in this paper provides an effective and safe way for the reliable preservation of RNA samples.Currently,commercialized room-temperature RNA storage kits,such as RNAshell?,RNAstable?,Integen X?,etc.,are available for RNA samples storage.This project evaluates the stability and effectiveness of samples preserved by RNAshell?,with a view to provide a simple,reliable,and economical method for RNA samples to be transported or stored at room temperature for short period of time.The project evaluates the stability of RNAshell?through accelerated degradation treatment?equivalent to storage at room temperature for 2 years,4 years,and 10 years?and real-time storage?4 years?,and statistically analyzes storage parameters including RNA recovery,purity and integrity.The results show that although the RIN value of RNAshell?samples has a tendency to decrease,its quality can still meet the requirements of downstream molecular biology research.The project also uses RNA-sequencing technique to further evaluate the impact of patented stabilizers in RNAshell?on high-throughput sequencing research.The results showed that the clean reads percentage of all samples have reached over 95%,with Q20>97%,Q30>92%and stable GC content.RNAshell?samples are safe to store with no significant impact on downstream high-throughput sequencing studies.It is worth mentioning that although RNAshell?has advantages mentioned above,its use requires corresponding supporting equipment to pre-treat the purified RNA samples,and the cost is relatively high.If RNA storage capacity of a biobank is lower than 10,000/year,then the cost of Cryopreservation will be lower than that of RNAshell?.While only if the RNA storage capacity of a biobank is between 100,000 and 1 million/year,and the RNA is kept continuously for 11 to 16 years,the cost of RNAshell?storage will show its advantages.To take cost-effectiveness into consideration and further brings down the cost for long-term preservation,we have configured 10 individual room-temperature stabilizers to test the protection effect of RNA samples of each experimental groups.The results showed that the freeze-drying state of room temperature stabilizer 2?1%SDS,2×RNAsecure and 20%trehalose?had an effective protective effect on RNA degradation?RIN value ranging from 9 to 7.4?,and the protective effect was closest to that of traditional ultra-low temperature preservation,which could be kept at room temperature for up to 28 days.Room temperature stabilizer2 for freeze-drying preservation method can be used as a cost-effective and reliable RNA preservation method,which can reduce preservation cost,guarantee sample quality and be environment-friendly at the same time.This project is of significant values in research and application for future RNA storage in biobanks. |
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