Nonradioisotopic telomeric repeat amplification protocol (TRAP) using digoxigenin labeled probe.
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.
Wada
Y
Y
Department of Surgery, Sapporo Medical University School of Medicine, Japan.
Yagihashi
A
A
Kameshima
H
H
Matsuura
A
A
Sato
N
N
Kikuchi
K
K
Yajima
T
T
Sasaki
K
K
Uehara
N
N
Watanabe
N
N
Hirata
K
K
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
D019098:Telomerase; D004076:Digoxigenin