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Given a set of paired-end reads R, maximizing the number of compatible reads with P is equivalent to maximizing), ∑ p ∈ R q p ~ P. For each gene, paired-end reads that are mapped to only one segment (i.e., | IS p| = 1) are excluded from R, since these reads do not provide any useful information in the assembly.
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The finding that 98% of gene fusions generate transcripts that can be translated as in-frame protein products confirms that reading frame conservation is of great functional importance in oncogenic fusion proteins, since the expected frequency of compatible reading frames between two random exons (assuming equal frequencies of +1, +2 and +3 reading frames) is one third.
Since all translocations in TICdb are mapped to specific introns or exons, we were able to check whether the exons flanking a translocation breakpoint have compatible reading frames, that is, if the last exon of the 5' partner gene ends in the same reading frame in which the first exon of the 3' partner gene starts.
Deletions in this exon always result in a compatible reading frame with the corresponding exons of FIP1L1 (exons 10, 11, 12 and 13 of FIP1L1 transcript ENST00000358575, which end in reading frames +3, +1, +2 and +3 respectively, in version 38.36 of Ensembl), leading to in-frame fusion transcripts in all four configurations.
Also, although the pipeline that was built around SPANNER was able to utilize the SR signal, its split alignment algorithm is only compatible with reads collected using the now defunct 454 sequencing technology.
Our protocol ensured that only those reads compatible with a gene model were used to evaluate the role of a genome annotation in RNA-Seq data analysis.
As SOAP2 is not compatible with intron-spanning reads we performed another mapping with the remaining unmapped reads with TOPHAT (version 1.3.1) to capture them as well.
The PSC8 samples infected with P. halstedii race 710 (a compatible interaction) generated 251,126 reads (with an average length of 176 bp and a median length of 161 bp), while the infected XRQ samples (an incompatible interaction) generated 161,526 reads (with an average length of 184 bp and a median length of 179 bp).
This new method does not require a ligation reaction, does not produce adapter dimers, does not require gel purification and is compatible with long sequencing reads the size of which is only limited by the length of library fragments and the sequencing technology.
Each read is processed as follows: If the isoforms compatible with the read correspond to nodes in more than one tree unite the corresponding trees.
EXPRSS Tag-seq was initially designed with Illumina single end adapters that are compatible with single read flow cells of GAII and Hiseq.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com