Sequencing potential of nested strand hybridization.

The capability of the sequencing by nested strand hybridization (SNSH) method to sequence unseparated pools of DNA fragments was assessed in computer simulation experiments. The results demonstrate the high resolving power of the method and its tolerance to false-positive errors. We determine the optimal proportion between the fragment length and the pool size at a given length of oligonucleotide probes, compare SNSH to the standard SBH, and suggest the best experimental setting for the special case of sequencing of long isolated fragments.