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This −34 position corresponds to −35 in our realigned sequence comparison.
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To overcome this problem, we realigned sequences within contigs using a custom script (Script available at: https://www.sourceforge.net/projects/snp454).net/projects/snp454
The intuitive graphical interface makes it easy to inspect, sort, delete, merge and realign sequences as part of the manual filtering process of large datasets.
For subsequent phylogenetic analysis, an HMM calculated for each alignment using hmmer [ 64] was used to realign sequences and to identify and remove indel regions and sequences with fewer than 60% matches to the model.
To further characterize these sequences we realigned our sequences with an extended HIV-1 A-like reference sequence dataset (including the subsubtype A3 which were not included in the original dataset but has been reported to circulate in West Africa, Table S3) using PRANK+F with a NJ tree (constructed in MEGA4) as guide tree [24], [46].
Groups from which sequences were removed due to excessive divergence are realigned after sequence removal with the same parameters before proceeding with analysis.
We realigned each sequence to the E. coli genome and clustered together the sequences that ended at the same 3' end.
For this, we identified related proteins from the multi-species neighbor-joining tree (as corroborated by dual Arabidopsis-other species trees), grouped closely related subgroups together, realigned these sequences, and inferred maximum likelihood phylogenies.
We substituted the corresponding sequence from ACI90286 for the run of Ns and realigned with our sequence data.
The realigned, recalibrated SAM (Sequence Alignment/Map) files produced by these processing steps were used for single-nucleotide polymorphism (SNP)/indel detection and for all alignment-related statistics, such as allele counts.
This would require realigning the sequences in the regions specified by "M" to retrieve the number of replacements used for alignment identity calculations.
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