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These extracted read pairs were then mapped to the Greengenes database using BWA-MEM v0.7.5a-r405 (Li and Durbin 2010).
In order to reconstruct breakpoint junctions, we sequenced FRAG00062 to 100× coverage, extracted read pairs from the ends of duplicated and triplicated blocks and performed de novo assembly.
Subsequently, we extracted read coverage at each genomic position and calculated, using R [ 39], the mean coverage of our reads on the genomes of wBm or B. malayi (Table 1, Table 2).
First, we extracted read depth information from the BAM files and compiled corresponding lists of mean read depths for each of the 30,521 genes defined by the gene models for the F. vesca 'Hawaii 4' v1.0 reference genome [ 41].
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Finally, the intrascaffold gaps were filled using the paired-end extracted reads.
The extracted reads were further clustered into nonredundant reads using USEARCH/UCLUST program v5.1 [ 67].
The extracted reads were de novo assembled to compile the genomic sequences flanking the inserted plasmid.
The extracted reads were assembled with Trinity using the parameters described above.
The extracted reads were taxonomically classified using the SILVAngs server (33).
For data analysis, we extracted reads starting with the 21-nt sequence xxxgaattcggccaggtacct.
We uniquely extracted reads covering heterozygous SNPs, from the BAM-file.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com