title: Nonradioisotopic telomeric repeat amplification protocol (TRAP) using digoxigenin labeled probe.
abstract: We have attempted to detect telomerase activity by Southern blot analysis using a digoxigenin labeled probe (5'-(CCCTTA)6CCCTAA-3') Telomerase activity was detected in all cancer cell lines that were tested, and:the lower limit for detection was 10(3) cells. Although telomerase activity was detected in colon cancer tissues, it was not found in adenoma or normal tissues. Telomerase activity was completely abrogated by heat and RNase treatment of the CHAPS extract. Our results demonstrate that this nonradioisotopic assay is safe and reliable for detecting telomerase activity in cancer cell lines and biopsy samples.
authors.affiliation: Department of Surgery, Sapporo Medical University School of Medicine, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Wada
authors.forename: A
authors.initials: A
authors.lastname: Yagihashi
authors.forename: H
authors.initials: H
authors.lastname: Kameshima
authors.forename: A
authors.initials: A
authors.lastname: Matsuura
authors.forename: N
authors.initials: N
authors.lastname: Sato
authors.forename: K
authors.initials: K
authors.lastname: Kikuchi
authors.forename: T
authors.initials: T
authors.lastname: Yajima
authors.forename: K
authors.initials: K
authors.lastname: Sasaki
authors.forename: N
authors.initials: N
authors.lastname: Uehara
authors.forename: N
authors.initials: N
authors.lastname: Watanabe
authors.forename: K
authors.initials: K
authors.lastname: Hirata
mesh_terms: D015139:Blotting, Southern; D004076:Digoxigenin; D005260:Female; D005784:Gene Amplification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012091:Repetitive Sequences, Nucleic Acid; D012680:Sensitivity and Specificity; D019098:Telomerase / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D019098:Telomerase; D004076:Digoxigenin