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Counting target molecules by exponential polymerase chain reaction: copy number of mitochondrial DNA in rat tissues

Item Type:Article
Title:Counting target molecules by exponential polymerase chain reaction: copy number of mitochondrial DNA in rat tissues
Creators Name:Wiesner, R.J., Ruegg, J.C. and Morano, I.
Abstract:In this report, we show that the actual number of target molecules of the polymerase chain reaction can be determined by measuring the concentration of product accumulating in consecutive cycles. The equation describing product accumulation, log Nn = log eff x n + log N0, can be analyzed by linear regression and the molar concentration of target at cycle zero, N0, is obtained. Using this new approach, the actual content of mitochondrial DNA was determined in rat tissues and ranged from 116 x 10(9) molecules/g in fast-twitch skeletal muscle to 743 x 10(9) molecules/g in liver. Using morphometric data from the literature, mitochondria were found to contain 1 to 3 DNA molecules. There was no relation between the oxidative capacity of a tissue and its content of mitochondrial DNA, indicating that transcriptional and posttranscriptional mechanisms rather than gene dosage, as postulated by others, determine to what extent the mitochondrial genome is expressed.
Keywords:Base Sequence, Mitochondrial DNA, Liver, Molecular Sequence Data, Multigene Family, Muscles, Polymerase Chain Reaction, Nucleic Acid Repetitive Sequences, Tissue Distribution, Animals, Rats
Source:Biochemical and Biophysical Research Communications
ISSN:0006-291X
Publisher:Academic Press
Volume:183
Number:2
Page Range:553-559
Date:16 March 1992
Official Publication:https://doi.org/10.1016/0006-291X(92)90517-O
PubMed:View item in PubMed

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