We sequenced the first 1000nt of the beta-actin gene from all twelve Lepomis species to generate primers that would identify DNA samples of each species and/or hybrids.

PCR products are species-specific.  Panel 3a shows the results when the “Exon 1 Forward Primer” is paired with the the Lepomis macrochirus Reverse Primer (MRP).  The expected product is 355 basepairs long.  Bands are only present with L. macrochirus DNA (lanes 2--4) and hybrid DNA (lanes 8--9).  Panel 3b shows the results when EX1F is paired with the the L. cyanellus Reverse Primer (CRP).  The expected product is 282 basepairs long.  Bands are only present with L. cyanellus DNA (lanes 5--7) and hybrid DNA (lanes 8--9).  Panel 3c shows the products of a reaction using EX1F, MRP and CRP.  Both species exhibit their expected size bands (lanes 2--4 and 5--7), and the hybrid DNA produces both 282 and 355 basepair products.

                            3a                                                            3b                                        3c