%0 Journal Article
%T The mitochondrial housekeeping gene 16S is inappropriate as an internal standard in comparative studies of rare mitochondrial transcripts using S1-nuclease protection assays
%A Sandra Ebert
%A Line Breumlund
%A Roland Nau
%A Uwe Michel
%J Journal of Nucleic Acids Investigation
%D 2010
%I PAGEPress Publications
%R 10.4081/jnai.2010.e2
%X The analysis of rare mitochondrial transcripts derived from the L-strand of the mitochondrial genome requires a sensitive method such as the S1-nuclease protection assay. We examined whether the ribosomal mitochon-drial transcript 16S is suitable as an internal standard in a multiplex S1-nuclease protection assay for the measurement of different mitochondrial transcripts. For reliable quantification of rare mitochondrial transcripts with the RNase protection assay, a minimum of 2 ¦Ìg of total RNA is necessary. Standard curves of 16S RNA produced with total RNA from human kidney, liver, brain, and a human neuroblastoma cell line (SH-SY5Y) revealed dose-response relationships that were saturated already at less than 0.5 ¦Ìg of total RNA. Therefore, 16S is inappropriate as an internal standard for analyzing mitochondrial transcripts with RNase protection assays when more than 0.5 ¦Ìg of total RNA have to be analyzed.
%K Rare mitochondrial transcripts
%K 16S
%K Housekeeping gene
%K Internal standard
%K S1-nuclease protection assay
%U http://www.pagepress.org/journals/index.php/jnai/article/view/1588