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Unlike other commercially available kits, exceptional quality and evenness of coverage are obtained even at very low levels of DNA input.
This eliminates many of the counting issues introduced by multiple tag approaches but reduces the evenness of coverage.
RNA and cDNA fragmentation are frequently used to improve evenness of coverage across the length of transcripts so splice junctions and SNPs can be more efficiently detected.
Reads are mapped to multiple alleles with limited mismatches, and the resulting allele candidates are compared statistically for the depth and evenness of coverage.
One fundamental problem is the lack of evenness of coverage [ 48], which is especially troublesome if the results are intended for diagnostic purposes.
Enzymes were also scored on evenness of coverage depth by counting the number of regions with zero and less than 5× base coverage.
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This strategy not only improves the evenness of read coverage but also can increase the coverage of the target genome (Rodrigue et al., 2009).
We evaluated the evenness of CDS coverage for each method, based on the L. lactis CDS coverage extracted from the MIX experiments.
We next determined the evenness of transcript coverage by comparing the median coefficient of variation (CV) for the read coverage of the 1000 most highly expressed transcripts (Table 1).
There was no correlation between the amount of input DNA, which ranged from 200 to 1000 ng in the pools, and the evenness of the coverage (Additional file 1: Figure S3), suggesting that using more of the wgaDNA would not have yielded more even coverage.
The success of a 454 sequencing strategy was evaluated by assessing the evenness of the coverage across the genome and the ability to assemble de novo the target genome.
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