He Xiao-Rui, Zhang Chunlian, Patterson Cam
University of North Carolina, Chapel Hill, NC 27599-7126, USA.
Biotechniques. 2004 Sep;37(3):464-8. doi: 10.2144/04373RT02.
A reproducible, transcriptionally diverse common reference RNA is required for accurate comparisons of data generated from most spotted microarray experiments in different experiments. Several methods have been proposed to make such a reference RNA, such as pooling RNAs isolated from multiple cell lines or tissues, amplifying pooled RNAs, or synthesizing RNAs or DNAs complementary to microarray features. We report an approach to prepare a large amount of mouse reference RNA from whole neonatal mice. This approach is simple, quick, reliable, reproducible, and inexpensive. The whole mouse reference RNA is highly representative when compared to two commercially available universal mouse reference RNAs isolated and pooled from multiple cell lines or organs.
在不同实验中,要对大多数点阵式微阵列实验产生的数据进行准确比较,就需要一种可重复、转录多样的通用参考RNA。已经提出了几种制备这种参考RNA的方法,比如汇集从多个细胞系或组织中分离得到的RNA、扩增汇集的RNA,或者合成与微阵列特征互补的RNA或DNA。我们报告了一种从新生小鼠整体制备大量小鼠参考RNA的方法。这种方法简单、快速、可靠、可重复且成本低廉。与从多个细胞系或器官中分离并汇集得到的两种市售通用小鼠参考RNA相比,整体小鼠参考RNA具有高度代表性。