Fig. 1. Polymerase chain reaction amplification of the epithelial SERCA cDNA fragment. (A) Amplification of a 513 bp band from total anterior sternal epithelium RNA using the degenerate primers SERCA-F1 and SERCA-R1; 5'-RACE (B,C) and 3'-RACE (D,E) were performed with the constructed adaptor-ligated cDNA pool to clone the full-length SERCA cDNA. The specific primers GSP1 and DAP-st11 (B) and the nested primers GSP2 and DAP (C) were used to amplify the 5'-end. GSP3 and DAP-TRsa (D) and the nested primers GSP4 and DAP (E) were used to amplify the 3'-end. The indicated bands at approx. 500 bp, 1.2 kbp and 2.6 kbp were extracted, purified, reamplified and sequenced. See Materials and methods for details.