Long-read sequencing (e.g. Nanopore's MinION or PacBio) has the potential to dramatically improve genome assembly but the quality of the reads is critically dependent upon the quality of the input gDNA. Here I describe the extraction of high molecular weight DNA from the calcareous sponge, Sycon capricorn. I choose to use Qiagen RLT buffer in the initial lysis step as this solution results in higher molecular weight DNA compared to alternatives. I suspect that RLT is deactivating endogenous DNases that would otherwise digest the gDNA.
According to the genetic rescue hypothesis, immigrants can improve population persistence through their genetic contribution alone. We investigate the potential for such rescue using small, inbred laboratory populations of the bean beetle (Callosobruchus maculatus). We ask how many migrants per generation (MPG) are needed to minimize the genetic component of extinction risk. During Phase 1, population size was made to fluctuate between 6 and 60 (for 10 generations). During this phase, we manipulated the number of MPG, replacing 0, 1, 3, or 5 females every generation with immigrant females. During Phase 2, we simply set an upper limit on population size (.10). Compared with the 0-MPG treatment, the other treatments were equivalently effective at improving reproductive success and reducing extinction risk. A single MPG was sufficient for genetic rescue, apparently because effective migration rate was inflated dramatically during generations when population size was small. An analysis of quasi-extinction suggests that replicate populations in the 1-MPG treatment benefited from initial purging of inbreeding depression. Populations in this treatment performed so well apparently because they received the dual benefit of purging followed by genetic infusion. Our results suggest the need for further evaluation of alternative schemes for genetic rescue.
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