RLM-RACE to determine transcription start sites in endometrial carcinoma cell lines. (A) Schematic outline of the RLM-RACE reaction consisting of the ligation of the 5'RACE-adaptor followed by 2 nested PCR reactions with outer and inner primers. (B and C) RNA preparations of Hec-1A, ECC-1, SPAC-1L, and Hec-1B cells were subjected to the RLM-RACE analysis. On the agarose gel the products of the nested PCR reactions obtained after outer (no product because of low copy number) and inner PCR are shown. Note the different size of amplified products. The water control served as a negative control without template. (D) Amplified bands were excised from the gel and subjected to DNA sequencing. The resulting products and the respective TSS are schematically indicated.