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Doing this, we selected, for each transcriptomic unit, transcripts that are preferentially over-expressed compared to the others.
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Depending on the analysis purpose, the probes were either grouped based on annotation units (such as exons) or transcription units (transcript isoforms).
The orientation of strand-specific sequencing reads matches the direction of transcription of a given transcription unit (for transcripts of genes found in the inverted repeat regions this becomes apparent only after the second copy of the repeat is removed from the reference sequence).
One of the major problems with tiling array data analysis is that the intensities of the multiple probes representing a given transcript unit (a transcript isoform) or annotation unit (such as an exon) are highly variable [27].
Because none of these methods was carried out at sufficient depth to identify all transcription units, evaluated transcripts were binned into three groups based on expression level (see Methods).
The read depth (i.e., the number of reads at a given position) was used to define a transcript unit and the transcript boundaries were manually annotated.
Of the reads that successfully hit the transcriptome those that were not aligned within 25 bp or more of the 3'- or 5'-end of the EST were considered gapped and were aligned against a collection of transcription units (Ensembl transcript, November 8th, 2005) using BLAT [ 18].
This was followed by extraction of meaning units from transcripts.
bThe coverage was calculated based on the unit of transcript.
The libraries contained an unequal number of initially used reads; therefore, comparisons in miRNA expression between the libraries were performed using the unit of transcript per million (TPM).
To describe transcript abundance across tissues, we mapped reads from our tissue-specific RNA-seq libraries to the AaegL3 genome and report transcript abundances in units of transcripts per million (TPM) [ 45].
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