Evaluation of methods for duration of preservation of RNA quality in rat liver used for transcriptome analysis
Evaluation of methods for duration of preservation of RNA quality in rat liver used for transcriptome analysis
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DOI：
10.2131/jts.31.509
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发表时间：
2006-12-01
影响因子：
2
通讯作者：
Urushidani, Tetsuro
中科院分区：
文献类型：
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作者：
Kasahara, Toshihiko;Miyazaki, Toshiko;Urushidani, Tetsuro
在毒理基因组学项目中，大约有150种化学物质被给予大鼠，肝脏中的基因表达由Affytron基因芯片分析并存储在数据库中。由于RNA的质量极大地影响基因表达数据的准确性，因此样品的储存条件非常重要。最近，一种RNA稳定化溶液，RNAlater（R），已经商业化。在这项研究中，新的存储方法与传统的存储方法（存储在冰箱或液氮中）在不同的条件下，通过观察其总产量，OD 260/280比，28 S/18 S比，和P-actin的数量评估的RNA的降解进行了比较。经证实，RNAlater（R）通过保持RNA的质量一年（在液体N-2或-80 ℃中）、3天（4 ℃）或2小时（室温）而不降解RNA来保存肝组织样品。溶解在缓冲液RLT中并储存在-20 ℃下的RNA样品的质量趋于下降，但储存在-80 ℃下的样品几乎与储存在液氮中的样品相当。综上所述，我们推荐以下程序保存用于提取RNA的肝组织：1）取出的组织应尽快放入冷冻的RNAlater（R）中; 2）RNAlater（R）中的样品必须在4 ℃下储存过夜或更长时间，并且可以在不冷冻的情况下放置长达2周; 3）在RNAlater中的样品可以在低于-20 ℃下保存至少一年，4）溶解在缓冲液RLT中的样品可以在-80 ℃下保存至少一年。
In The Toxicogenomics Project, about 150 chemicals are administered to rats, and gene expression in the liver analyzed by Affymetrix GeneChip and stored in the database. As the quality of RNA greatly influences the accuracy of gene expression data, conditions of the storage of the sample are very important. Recently, an RNA stabilization solution, RNAlater (R), has become commercially available. In this study, the new storage method was compared with the traditional storage method (stored in freezer or liquid nitrogen) under various conditions by looking at the degradation of RNA assessed by its total yield, OD260/280 ratio, 28S/18S, ratio, and quantity of P-actin. It was confirmed that RNAlater (R) preserved the liver tissue sample by maintaining the quality of RNA for one year (in liquid N-2 or -80 degrees C), for 3 days (4 degrees C), or for 2 hr (room temperature) without degradation of RNA. Quality of RNA samples dissolved in buffer RLT and stored at -20 degrees C tended to decrease, but samples stored at -80 degrees C were almost equivalent to those stored in liquid nitrogen. In conclusion, we recommend the following procedure for preservation of liver tissue for extraction of RNA: 1) tissues removed should be put into chilled RNAlater (R) as soon as possible; 2) samples in RNAlater (R) must be stored overnight or longer at 4 degrees C and can be left for as long as 2 weeks without freezing; 3) samples in RNAlater can be stored for at least one year under less than -20 degrees C and 4) samples dissolved in buffer RLT can be preserved at least for one year under -80 degrees C.