Because of DNA sample limitations, we utilized only a subset of the previously studied patient cohort.
Despite the reliability and validity found for pooled DNA, three limitations of this study should be mentioned, which put these results in an even more favourable light.
Robustness is a remarkable feature of living organisms allowing them to tolerate a wide variety of contingencies, such as DNA damage, limitations in nutrient availability, or exposure to toxins (Lopez-Maury et al., 2008; MacNeil and Walhout, 2011).
However, without DNA resource limitations, a genotyping bottleneck exists mostly as a result of time- and assay set-up costs and hence, in order to achieve large-scale genotyping, highly multiplexed assays are required.
The regions encoding BEX2 with 3 CpG sites and cXorf23 with a single CpG site were each run in a single sample well due to DNA quantity limitations (N = 59 including the original 46 profiled on BeadChip).
While these high-throughput methods are powerful, there are practical limitations: DNA microarrays are limited to detecting genes for which allelic variants have already been characterized and may miss emerging mutations; the PAML-based approach requires multiple genomes of phenotypic variants of the same species, which are not always available.
However, an important limitation of DNA vaccines is limited potency since they lack the intrinsic ability to amplify and spread in vivo.
To cope with such limitations, DNA barcode sequences have been analysed based on other species concepts [38], and referred to as Recognizable Taxonomic Units [43], or Molecular Operational Taxonomic Units [44].
However, DNA microarrays have limitations including background hybridization and known probe information.
Finally, bisulfite conversion and pyrosequencing as a method of determining DNA methylation has limitations, including the possibility of incomplete bisulfite conversion and biased amplification.
The chemical synthesis of DNA molecule has limitations in terms of strand length; thus, the creation of artificial genes requires the assembly of long DNA molecules from shorter fragments.
