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Quantitative analysis was performed using CLCbio (CLC Genomics Workbench 4.8, parameter settings: minimum length fraction, 0.5; minimum similarity fraction, 0.95; maximum number of hits for a read, 10).
The following criteria were used to filter the unique sequence reads: minimum length fraction of 0.9; minimum similarity fraction of 0.8; maximum number of two mismatches.
The RNA-seq analysis tool included in the CLC Genomic Workbench was used for this purpose (minimum length fraction and minimum similarity fraction were set at 0.75 and 0.95, respectively).
The "RNA-Seq analysis" option was used to map reads to each database at the following settings: minimum length fraction 0.9, minimum similarity fraction 0.8, and maximum number of hits for a read 30.
Alignment settings for color space reads were the following: maximum number of mismatches: 2; minimum length fraction: 0.9; minimum similarity fraction: 0.8 and maximum number of hits for a read: 10 (see Additional file 13A for mapping statistics).
The alignment parameters were adjusted to allow for expected variations in the octoploid genome relative to the diploid one (Minimum length fraction = 0.6; Minimum similarity fraction = 0.5; Maximum number of hits for a read = 5).
Similar(48)
To obtain the gene expression values, reads from each sample replicate were mapped to the genes (i.e. assembly contigs) with the default mapping parameters {minimum similarity fraction = 0.8 and minimum length fraction (long reads) = 0.9} [ 75].
The maximum number of mismatches allowed was two; the minimum length and similarity fraction was set at 0.8; and the maximum number of hits per read was 10.
The sequencing reads from each tissue were mapped back to the reference unigenes based upon a minimum similarity of 0.9 and length fraction of 0.5 [ 66].
CLC assemblies were conducted using the following parameters: mismatch cost = 3, insertion cost = 3, deletion cost = 3, length fraction = 0.5, similarity fraction = 0.8, minimum contig length = 200, automatic bubble size = yes, automatic word size = yes, perform scaffolding = yes.
The parameters used for de novo assembly are: Mapping mode = Map reads back to contigs (slow); Similarity fraction = 0.8; Minimum contig length = 200; Mismatch cost = 2; Insertion cost = 3; Deletion cost = 3; Update contigs = Yes; Length fraction = 0.5; Perform scaffolding = Yes.
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