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To screen the Violaceae alignment for possible recombination we employed the Genetic Algorithms for Recombination Detection (GARD) [ 46, 47], on the exons only, using general parameter settings (GTR model of nucleotide substitution and beta-gamma rate variation with 2 rate classes).
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For every sequence family in BAliBASE 3.0, we performed pairwise sequence alignment for all possible sequence pairs in the given family.
We obtained multiple sequence alignments for all possible protein isoform combinations in each of the families of the four data sets (table 2) using MAFFT with default parameters (Katoh et al. 2002; Katoh and Toh 2008).
Sequence alignments were cleaned for possible misaligned sites running Gblocks with default parameter settings [ 57].
Sequence alignments were scanned for possible recombination using the software package RDP3, employs a suite of recombination detection and analysis methods [ 67].
In some instances, all sequences within alignments were screened for possible signal peptides using SignalP v.4.0 (Petersen et al. 2011), LipoP v.1.0 (Juncker et al. 2003), and Phobius (Kall et al. 2007).
An alignment for all 17 possible medium-chain dehydrogenases/reductases (MDR) enzymes of E. coli revealed the closest relationship of YahK and YjgB to each other without mentioning YqhD (Jörnvall et al. 1999b).
Reads were independently aligned to the S.erythraea genome using multiple software to assess for possible alignment bias caused by the high GC content of the genome.
Every location is then refined using an optimal alignment algorithm specifically accounting for possible splice sites.
We aligned the amino acid sequences of each pair of orthologs from nine- and three-spined sticklebacks using MUSCLE [ 77] with default settings and manually inspected for possible alignment artifacts.
Alignment only possible for small area (from mm2 to cm2).
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