A 20 l RTreaction was performed with the Expand RT-PCR Kit from
Then 2.5 l of the RT reaction were
employed for PCR, using the Taq Polymerase from Invitrogen
(Karlsruhe, Germany) with CYTGAAYSASCGYYTGGC as degenerate
forward and GTAKGTBGCWATYTCMRCHTCSAG as degenerate
reverse primer; we applied a standard three step program (5 min
initial denaturation at 94C; denaturation for 40 s at 94C; annealing for
30 s at 47C, elongation for 45 s at 72C; final extension for 7 min at
72 C)