The adaptor-ligated DNA was hybridized for 64 h at 42 [degrees]C to one of 4 independent array sectors for samples CPSA, CPSB, and OTCB with hybridization conditions that were slightly modified compared with those for sample OTCA (see Methods in the online Data Supplement).
A 454-sequencing service provider (Microsynth) prepared a single sequencing library from the tagged and equally pooled samples CPSA, CPSB, OTCB, and UNC, and a separate library for sample OTCA.
For the pooled library of samples CPSA, CPSB, OTCB, and UNC, we obtained a total of 584 390 sequence reads and 181 Mb of sequence with a mean read length of 311 bp.
This result indicated a more homogeneous sequence distribution across the target region for samples CPSA, CPSB, and OTCB, potentially because of the slightly modified hybridization protocol (Table 1).
The 3 samples captured with an identical protocol (CPSA, CPSB, and OTCB) had very similar distributions of sequencing depth, which demonstrated the high intersample reproducibility of the MSC procedure (Table 2, Fig.