Additional file 2.

Sequence data and PCR conditions for specific primer combinations used for all six species of Poeciliidae. Primer names are given with corresponding primer number from Additional file 1. Amplicons are shown in Additional file 3 along with corresponding n value. 0.5 U iProof™ DNA polymerase (BioRad®) was used for each reaction with 5× iProof™ HF Buffer, 10 mM dNTP mix, ~100 ng template DNA, 0.5 μM of both forward and reverse primers and dH2O. An Eppendorf® silver block thermal cycler was used for all PCR reactions. Each reaction included an initial denaturation of 94°C for 30 seconds and a final extension of 72°C for 600 seconds. If >1 bands were found after gel electrophoresis, expected sized amplicons were cut and purified using a QIAquick® Gel Extraction Kit.

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Ward et al. BMC Evolutionary Biology 2008 8:210   doi:10.1186/1471-2148-8-210