Unless the environment has changed drastically since a population became fragmented and the population fragments have become locally adapted, outbreeding depression is not expected to be a problem at this scale as increased connectivity only restores the previous situation.
The genetic effect of habitat fragmentation depends largely on the level of gene flow within and among population fragments.
First, the variation can be replenished by gene flow between population fragments.
Improving gene flow and connectivity between the population fragments that previously formed a single large population is the first step.
By contrast, the 454 library preparation involves random shearing of the normalized but un-cloned cDNA population, fragment end polishing, adaptor ligation, library immobilization, fill-in reaction, single stranded DNA library isolation and pyrosequencing [ 2].
The lymphocyte subpopulation was gated according to standard Forward and Side Scatter parameters, in order to avoid irrelevant larger cell populations or smaller cell populations, fragments and debris.
This technique can be used by wildlife managers and road planners, and may be particularly important in reduction of road mortality for species such as the Iberian lynx (Lynx pardinus), Florida panther (Puma concolor coryi), and Texas ocelot (Leopardis pardalus), which exist in small populations fragmented by development, and are limited by road mortalities.
However, some DNA-protein interactions may remain undetected because of low affinity or the low concentration of an individual fragment in a large population of fragments [ 11].
Previously, two similar high-throughput methods were developed for the identification of protein binding regions using a large population of fragments derived from DNAs (plasmids, bacteriophages, bacterial chromosome and human genome fragment) ranging in size from 3 kb to 4,700 kb [ 10, 11].
An extension of this method uses the large population of fragments derived from long genomic regions (approximately 600 kb) for the identification of fragments containing protein binding regions.
However, this strategy has the drawback of producing a population of fragments smaller than 2 Kb; as a result, the fragmentation process used in the standard 454 GS FLX (Roche) shotgun procedure (nebulization), may not give optimal results since it is more suitable for high molecular weight molecules.
