Hybridization stringencies were adjusted to accommodate this degree of identity for all of the comparative data described in this study.
The soil metagenomic DNA was targeted at four different probe hybridization stringencies and sequenced with high-throughput sequencing.
Lower hybridization stringencies also increase the fraction of proteins without any annotations in the public databases in the metagenome.
It could also be inferred that this technique is reproducible as there is an increase in target enrichment while maintaining diversity at higher hybridization stringencies for both metagenomes.
The microarray includes 60 negative control probes and six mismatch hybridization stringency control probes designed against each of 60 contigs with the highest EST count in the database.
After hybridization, stringency washes were done with 4× SSC, 2× SSC–50% formamide at 45°C, and again with 1× SSC at room temperature for 30 minutes.
After overnight hybridization, slides were washed in 20% (v/v) formamide/0.1 × SSC at 42°C, giving a hybridization stringency of 85%.
The results of the CDS presence/absence analysis were, however, related to the high hybridization stringency conditions used, which may not allow detection of barely similar homologs.
The functional subsystems distribution between the captured metagenomes was mainly influenced by their soil type rather than their hybridization stringency, although both properties significantly affected their distribution.
A "touchdown" PCR cycling regimen and the addition of dimethyl sulfoxide (10% final v:v) was used in order to optimize the hybridization stringency.
