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For this reason, the search for novel and known miRNAs in our ZF samples was complemented by a megaBlast alignment between our dataset and mature sequences deposited in miRBase 12.0 and also the novel miRNA transcripts predicted by Ensembl and by Thatcher et al [ 44] (see Additional File 1).
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These A. marginale sequencing reads were then used for a second Megablast alignment using as a reference sequence 28 nucleotides from the Himar1 terminal inverted repeats (TIR).
The derived cDNA sequences were compared by megablast alignment with genomic contigs of an early version of the D. labrax whole genome shotgun assembly (Max Planck Institute for Molecular Genetics, Berlin, Germany).
Every read was mapped to a single ORF of the highest bit score if the MegaBLAST alignment had an identity ≥90% and e-value ≤10−5.
St Maries reference genome sequence (CP000030) using the Megablast alignment tool (NCBI BLAST + blastn (version 0.0.12) in GALAXY) to obtain sequencing reads that contained A. marginale sequences.
Optimization of word count for MegaBLAST alignment to the NT reference database.
- = no significant MegaBLAST alignment with Os_CD database Figure 3 illustrates that the MWs based upon percentage numbers of alignments generally coincide with MWs based upon average scores.
Sequences that were >99.5% identical over >90% of the ITS region to more than one type sequence identified in GenBank or RefS/RepS from UNITE of a different TaxID using megablast alignments.
A fully valid reporter must have MegaBLAST alignments to the RefSeq RNA transcripts of exactly one gene in Entrez Gene, and at least one of those alignments must be high-quality as defined in Methods.
BACs were masked for repeats and MegaBLAST alignments of these BACs were performed against a database of WGS reads.
We then performed MegaBLAST alignments of the WGS reads to the reference genome.
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