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We chose to measure mRNA of one gene from each transcript of the two strands of the D-loop that control replication and transcription of mtDNA, in order to detect changes in mitochondrial-encoded gene activity.
For each transcript of interest, known amounts of plasmids with this transcript were used to create a standard curve.
All samples were run in quadruplicates and in addition four independent parallels were used to generate the data for each transcript of interest.
Standard deviation for expression level of each transcript of interest was calculated for each condition.
Fluorescence readings from blank wells were subtracted from the fluorescence values for each transcript of interest.
Each sample was measured in triplicate for each transcript of interest and an internal reference gene.
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In addition, percent composition of each transcript component of each unigene was calculated.
Intriguingly, each transcript isoform of Hel1-333 was unique in each inbred and displayed different combinations of splice site usage.
Each transcript variants of Rae-1δ and Rae-1ε are differentially expressed according to tissues, pathological conditions and cell lines.
For each transcript, positions of introns were inferred from genomic intervals between consecutive exons.
Having transcript measures from different tissues from the same individuals allowed us to evaluate, for each transcript, sources of transcript level variation within and between individuals (Fig. 1).
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