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All are acceptable, although there was consensus that the SSU rRNA gene (SSU rDNA) was the optimum locus for genotyping for several reasons.
For subtyping, the second internal transcribed spacer region of nuclear ribosomal DNA (ITS-2) (GenBank accession no. AF015774) was considered the optimum locus and is thought to be sufficiently variable as well as being present in multiple copy number (five).
If choices were to be made as to which method to apply in any particular circumstance, the SSU rDNA was considered the optimum locus to target for reasons that included the size of the current database and the availability of primers that would detect the presence of all known genotypes.
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For unknown reasons, heavily inflated maps with a large number of misplaced loci were often reported by JoinMap to be shorter than maps constructed from the same datasets using JoinMap settings that produced a close-to-optimum locus order.
Where there were choices as to which method to apply, the small-subunit ribosomal RNA gene was considered to be the optimum genetic locus to target.
The spreadsheet produced by MSATCOMMANDER was used to select a small subset of 63 loci with optimum primer conditions and loci that had no lowercase letters in the primer sequence that would indicate nucleotide mismatches in the assembled reads used to create the contigs.
Agreement now on the optimum or consensus locus would help future researchers compare future with past results.
This strategy differs from the alternative approach of constructing a consensus map using the segregation data from different mapping populations to compute the optimum order of loci [ 32].
In strategy I (used in this study) the segregation data from several populations are simultaneously considered to compute the optimum order of loci.
A Kosambi map function was used to convert the recombination frequency into cM distances and the third round was set to allow the mapping of an optimum number of loci in the genetic map.
Fifty loci with optimum primer conditions were screened for amplification success and within-population variability.
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