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We also used the 27 species multiple alignment available from the UCSC genome center (http://genome.ucsc.edu/) to orient variation.
Then, for the homologous regions scanned for polymorphism, we compared the human, chimpanzee and macaque sequences in the 27 species multiple alignment available from the UCSC genome center to get fixed indel and single nucleotide differences between human and chimpanzee.
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Orthologous promoter regions were extracted from multiple alignments available at the UCSC Genome Browser (http://genome.ucsc.edu) (Karolchik et al. 2003; Karolchik et al. 2007), using known RefSeq TSS annotations in humans (Maglott et al. 2000; Pruitt et al. 2000).
We retrieved the Pecan 10 amniotes multiple alignments available at the Ensembl database (version 56) corresponding to each window and restricted these to the analysis of the following species: human, chimpanzee, and macaque for the analysis of the human lineage, mouse, rat, and human for the analysis of the mouse lineage.
As can be seen on the multiple sequence alignment (available in TreeBASE), we could not align the 14 blocks with enough confidence (p < 0.01) on the corresponding query sequences, the worst case being the N. capensis ω virus sequence for which only 3 blocks could be confidently aligned.
At the completion of each stage, a multiple alignment is available and the algorithm can be terminated.
Specific primers for each family of GPCR Class A were designed using conserved gene regions identified by multiple alignment of available olfactory receptors from teleost fish species.
Although ChimeraSlayer is presented as a 16S-specific method, it would likely perform well with another sequence type if a reference multiple alignment is available.
Multiple alignment of available GTPase sequences was accomplished using a previously described strategy [ 11] that iterates between sequence detection and alignment using the MAPGAPS program [ 31] http://mapgaps.igs.umaryland.edu.edu
The RFam database (Gardner et al., 2011) contains a high-quality set of ncRNA families, but its scope is limited to families for which an expert multiple alignment is available.
Sequences were aligned with Clustal W1.75 program (17 ) into MSF-format and edited with GeneDoc Multiple Sequence Alignment (available from: http://www.psc.edu/biomed/genedoc/) editor program.
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