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There are 926 codons (2778 bp) in the alignment.
We then selected from 1 3 pairs of CNE from each linkage group, as long as they satisfied the following criteria: i. highly similar sequences over at least 25 30 bp in the alignment between fugu-zebrafish-human; ii. a physical distance of more than 1000 bp and less than 10 Kbp between the two CNE with a conserved association in all species; and iii.
Similar(58)
Gaps of > 200 bp in the alignments were defined as CNVs and annotated as insertions or deletions with respect to the reference genome.
In addition to polymorphism clusters, we identified moderately sized indels (30 50 bp) in the alignments and selected 80 bp of total sequence for each species including the insertion when present and flanking sequence centered on the middle of the indel.
A cutoff value of 90% identity and 60 bp length in the alignment between the candidate homeologs and the reference sequence were used.
The resultant alignment was then checked using JalView ver. 8.0 [62], totaling 595 bp in the final alignment (File S2).
The liver-expression-specific nucleotide closest to the TSS is a C at the 3028th bp in the multiple alignment of 3447 bp of the putative promoters for the six UGT1A isoforms with hepatic expression and either a gap or a G for the three UGT1A isoforms not expressed in liver (see supplemental materials).
When there were gaps or short inserts (1-10 bp) in the selected alignment region then the corresponding sequences were checked using the Ensembl Genome Browser.
The rate of the five combined mitochondrial markers (cox1, rrnL, trnL, nad1 and cob, comprising 2,293 bp in the MF alignment; Table 2) was 0.010 +/- 0.002, equivalent to an overall pair wise divergence of 2.0% per MY (Table 3).
We calculated average read depth per segment of 100 bp in the mtDNA alignments both before and after filtering and compared the two sets of read depths.
This resulted in several assembled contigs that were just under 200 bp (but most were between 400 and 600 bp) to be included in the alignment.
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