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18–20 W). 3. Heat denature the CFLP reactions at 80°C for 2 min immediately prior to loading onto the gel. The best resolution is achieved when the samples are fully denatured. 4. Load 5–10 µL of the appropriate CFLP reaction per well. The remainder of the reactions can be stored at 4°C or –20°C for later analysis. 5. Continue electrophoresis until sufficient separation of the CFLP fragments is achieved (the time will depend on the fragment size and the percentage of acrylamide used). 4. 1. Fluorescence Imaging of CFLP Patterns 1.

Holt 1. Introduction The onset of human cancer typically requires numerous genetic mutations, generally specific for the tissue type from which the cancer originates. Thus, it has been difficult to screen all tumor types for a single mutation. In recent years, telomerase activity has been associated with at least 85% of human malignancies as well as with some lesions considered preneoplastic by traditional cytology (1,2). Telomerase appears to be ubiquitously associated with a wide array of human cancers from a variety of tissue sources.

3. 4. 5. 6. 7. 8. For step 2, make up 100 mL of 50% ethanol + 10% acetic acid in deionized water. For steps 3, 4, 6, and 7, make up a total of 500 mL of 10% ethanol + 5% acetic acid. For steps 8, 9, 11, and 12, set aside 1000 mL of deionized water. For steps 13 and 14 (developer), pipet one ampoule of 2% formaldehyde into one bottle of sodium carbonate (both of these are provided in the kit). Mix thoroughly. For step 15, prepare the background reducer by adding one packet of sodium thiosulfate into 100 mL of deionized water.

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