DNA Extraction From Sofkahatchee Stippled Studfish

This entry's title is more of a mouthful than I intended. But, yesterday Travis and I extracted DNA from four of the bifax from Sofkahatchee Creek that Joe Scanlan caught and gave to me. It was Travis' first extraction, using the old-fashioned PCI/CI phase separation method with white muscle tissue. Everything seemed to go well. We didn't get big pellets at the end, which is usually a good sign since pellets contain more protein and salts than DNA, and in truth I could see just indistinct smears along the tubes' walls. Leland graciously agreed to help us used the Nano Drop spec in the Podila lab, and we measured DNA concentration values of 30-90 nanograms/microliter with 260/280 ratios of 1.6-1.8. I was a little put off by these values at first, since our first round of extractions had concentrations more in the range of 100-300 ng/ul and ratios of 1.8-2.1. But Leland wasn't phased, saying that was their typical values for PCR work in their lab. Maybe we just did a better, cleaner job of extracting DNA with few contaminants? I guess so; or the Nano Drop wasn't as clean before, which Leland said was also the case? We'll know once we run PCRs on these samples. We haven't been able to assay concentrations for a while, and probably our most recent extractions have the same values as yesterday's and produced good bands on gels after PCR. I guess the real proof will be looking at sequences, hopefully sooner than later.