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The mapped partner of a MUR may contain mismatches and may have multiple mapping positions across the whole genome.
The original reads were first aligned onto the Sanger-obtained sequence of the region by using the same parameters described above and by allowing random matches of reads with multiple mapping positions.
When multiple mapping positions were obtained for the same read, a single hit was considered (the most significant one).
Filtered reads of all three replicates in each species were mapped to the corresponding pre-miRNAs in the same gene family using Bowtie 0.12.7, which only allowed at most 13 multiple mapping positions (the largest number of miRNA gene copies in two species, table 1) and zero mismatch for each read (Langmead et al. 2009).
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Therefore we included piRNAs that mapped to multiple positions in the genome, but divided the number of such multiple-mapping piRNA reads by the number of mapping positions when estimating their relative abundance (e.g., a piRNA mapped to 10 positions was counted as 0.1 reads at each position).
When calculating the number of piRNAs overlapping MEs in the genome, piRNAs that mapped to multiple locations were divided by the total number of mapping positions.
Multiple position mapping was enabled and the weight of multiple mapping reads was considered.
Besides that, we define Single Best Match mappings as multiple mapped reads with a unique best match position in the reference sequence.
Jiang and Pugh developed a compiled reference map of nucleosome positions in Saccharomyces cerevisiae, using multiple maps independent of our study (Jiang and Pugh 2009a, b).
Short reads are also aligned following less stringent criteria allowing reads to map multiple positions, which helped in identification of repeat genes.
Superscaffolds that were anchored in multiple maps were used as reference points to align the genetic positions in the three different maps.
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