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Mapped assemblies were converted to a gap5 database [ 47].
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Hybrid mapping assemblies were generated using Newbler reference assembler (120 contigs) [19], or CLC Genomics Workbench (3 contigs; CLC bio, Cambridge, MA) reference assembler and are summarized in Table 1.
However, mapping assemblies were best performed by CLC Genomics Workbench.
Except for the first WGP assembly, all physical map assemblies were performed using the methodology described by Paux et al [ 23] for the construction of the physical map of chromosome 3B.
In addition to demonstrating that residual heterozygosity is limited in the inbred lines, this analysis suggests that, in general, the expectation that homeolog copies will tend co-assemble in the reference transcript assembly and subsequent mapping assemblies is valid.
The size of the physical map assemblies was consistent with the relative sizes of each chromosome [17].
Details of the procedure are described in the Methods section and a summary of the successive physical map assemblies is given in Table 2. aAn initial test to determine the optimal cutoff value was performed with several cutoff values in the 1e-35/1e-65 range bNumber of contigs containing ≤/> than 5% of Q-clones cA cutoff value of 1e-45 was used for the initial contig assembly.
The influence of DNA molecule depth on map assembly were investigated and discussed.
The QC module was then used to perform a QC review to ensure all regions of map assembly were robust.
The physical map assembly was validated at several scales.
The reliability of the map assembly was validated by PCR assays on randomly selected 21 contigs.
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