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Only reads that aligned to a unique location in the genome were used to calculate the expression levels.
To identify genes with different expression levels, we used the RPKM method (Reads Per kb per Million reads) to calculate the expression levels of the unigenes.
To calculate the expression levels of CEP genes, RNA-Seq reads were remapped to their reference genomes using the short-read aligner SMALT [ 75] with default settings.
To calculate the expression levels of transcripts, we only keep probesets whose measured exons correspond to only one RefSeq transcript (called "unique" probesets).
The RNA-seq mapping performed to calculate the expression levels of the assembled transcripts in both analyzed organs mapped the majority of paired-end reads (Table 5).
Small RNA 202 was used as control to calculate the expression levels of two microRNAs, miR-34a and -196a in isolated biological specimens.
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Reads that could be uniquely mapped to a gene were used to calculate the expression level.
To calculate the expression level of each gene, we referred to the methods from the previous work [ 9].
For genes having more than one transcript, the longest one was used to calculate the expression level.
All samples were amplified in triplicate and the mean values were used to calculate the expression level of each target.
Different summation methods to calculate the expression level for each miRNA transcript from the individual isomir complement have been proposed.
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