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The multiple k-mer assemblies encompass both higher k-mer lengths to result in a more contiguous assembly of highly expressed transcripts, and lower k-mer lengths to better assemble poorly expressed transcripts to increase assembly sensitivity.
A higher k-mer length will theoretically results in a more contiguous assembly of highly expressed transcripts.
The sequences can be aggressively merged creating false joins in some cases but at the same time producing a statistically more contiguous assembly.
So, a higher k-mer length will theoretically generate a more contiguous assembly of highly expressed RNAs while poorly expressed transcripts will be more easily obtained if a lower k-mer length is used.
While such filtering may have biased our data set towards more highly expressed genes, it also yielded a more contiguous assembly, as the percentages of sequences larger than 1000 bp were doubled and the percentages of the short sequences were reduced by half (Table 2).
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The use of high K-mer lengths can generate more contiguous assemblies of highly expressed transcripts.
As a rule of thumb, large genomes have longer genes and thus need more contiguous assemblies for successful annotation (cf. Figure 1 in Yandell and Ence 2012).
Therefore, although our data clearly demonstrate the key role homology unit duplication has had in evolution of the bovine TRAV/TRDV repertoire, analysis of improved, more contiguous assemblies of the bovine TRA/TRD locus will be required to fully appreciate the extent and complexity of homology unit duplication.
These results are consistent with improvements in the updated assembly of v4 and with GreenGenie2 providing reliable predictions on a more contiguous genome assembly.
Previous studies showed that de novo assembly using the Velvet program with longer k-mers results in a more contiguous transcript assembly but lower transcript diversity compared to shorter k-mers [ 32, 33].
Divergence from the organization of the Nicotiana reference likely prevented a more complete and contiguous assembly.
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