Illustra ready to go pcr beads

Genomic DNA was extracted from liver samples following the protocol used by Medeiros et al. [17 (link)]. The H1 and cytb fragments were amplified by PCR using the Illustra Ready-To-Go PCR Beads (GE Healthcare, Chicago, IL, USA), the primers MVZ59/MVZ50 [27 (link)] and 12SL13 [28 (link)]/16Sbr [25 ] in the case of the H1 fragments, and the primers MVZ15 [29 (link)] and H15149(H) [30 (link)] for the amplification of the cytb fragments. The resulting amplicons were purified using the Wizard SV Gel and PCR Clean-Up System (Promega, USA), and sequenced with the same primers used for PCR. In the case of the amplicon obtained with the primers 12SL13 and 16Sbr, the following primers were also employed for sequencing: H1KR [31 ], Hedges16SL2a [32 (link)], Hedges16SH10 [32 (link)], and 16Sar [25 ]. Sequencing reactions were prepared using BigDye Terminator (Applied Biosystems, USA), according to the manufacturer’s instructions. Nucleotide sequences were obtained by an ABI PRISM 3130XL Genetic Analyzer automated sequencer (Hitachi, Tokyo, Japan) at the DNA Sequencing Facility of the Chemistry Institute of University of São Paulo.