Detection of microsatellite instability in cancers by arbitrarily primed-PCR fingerprinting using a fluorescently labeled primer (FAP-PCR).
The microsatellite mutator phenotype (MMP), detected as a change in the number of repeating units in hundreds of thousands of microsatellite sequences in the tumor cell genome, underlies the carcinogenesis of a variety of tumors including sporadic and hereditary nonpolyposis colon cancers. This enhanced microsatellite instability was discovered using arbitrarily primed polymerase chain reaction (AP-PCR) fingerprinting of DNA from colon cancers. In this study, we found an arbitrary primer that can amplify multiple DNA fragments containing repeated sequences, including the poly A tracts found in the Alu repeats of the human genome. The combined use of primer labeling with fluorescence and an automated DNA sequencing analysis of AP-PCR products (FAP-PCR) detected alterations in fingerprint bands in all DNA samples previously determined to belong to the MMP. Fluorescent AP-PCR fingerprinting using this single arbitrary primer provides a convenient and efficient method for detecting tumor specific fingerprint alterations that are usually undetectable by conventional fingerprinting.
Yasuda
J
J
Oncogene Division, National Cancer Center Research Institute, Tokyo, Japan.
Kashiwabara
H
H
Kawakami
K
K
Uematsu
K
K
Sugano
K
K
Perucho
M
M
Sekiya
T
T
D015139:Blotting, Southern; D002877:Chromosomes, Human / Q000737:chemistry; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000032:analysis; D005456:Fluorescent Dyes; D006801:Humans; D018895:Microsatellite Repeats; D009369:Neoplasms / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004273:DNA, Neoplasm; D005456:Fluorescent Dyes