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DNA sequence alignment, coding region assembly, and in silico translation were done using the DNAsis Max program 2.0 (Hitachi).
Annotation of de novo assembled contigs was done by alignment coding sequences against the human RefSeq mRNA database (Homo Sapiens GRCh37.66.cdna; http://www.ensembl.org/Homo_sapiens).org/Homo_sapiens
Three data sets were analyzed: those portions of the NA alignment coding for the ThiF domains of the MoeZ conceptual proteins, the alignment of the complete MoeZ sequences, and the alignment of the full length 16S rDNA (SSU) sequences.
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In more distant species, this ratio suddenly reverses, and in human- Danio rerio alignments, coding sequences are more conserved than non-coding (CNC: mean 38.58 % identity and CDS: 62.77%) (see Figure 4A).
6 Multiple alignment of coding DNA from aligned amino acid sequences was done using RevTrans.
In fact, some of the code written for Confab has been merged into the main Open Babel distribution (such as the original Kabsch alignment code) but due to an additional dependency (on tree.hh, see below) the core code has not been included in Open Babel v2.3.
All alignment code was run on the NIH Biowulf supercomputing cluster.
An alignment of coding sequences corresponding to the protein alignment was constructed using the tranalign program from the EMBOSS package [ 34].
For the alignment of coding sequence, sequences were translated, aligned at the amino acid level, and then back-translated to produce alignments at the DNA level.
All data sets were then converted to codon alignments using the University of Bergen computational biology unit alignment to coding tool (www.bioinfo.no).
This method computes the alignment of coding nucleotide sequences with respect to their possible translations.
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