학술논문

Detection of point mutations by non-isotopic single strand conformation polymorphism
Document Type
other
Source
Brazilian Journal of Medical and Biological Research. January 1999 32(1)
Subject
p53 gene
human papillomavirus
PCR
SSCP
cancer
silver staining
Language
English
ISSN
0100-879X
Abstract
We have developed a procedure for nonradioactive single strand conformation polymorphism analysis and applied it to the detection of point mutations in the human tumor suppressor gene p53. The protocol does not require any particular facilities or equipment, such as radioactive handling, large gel units for sequencing, or a semiautomated electrophoresis system. This technique consists of amplification of DNA fragments by PCR with specific oligonucleotide primers, denaturation, and electrophoresis on small neutral polyacrylamide gels, followed by silver staining. The sensitivity of this procedure is comparable to other described techniques and the method is easy to perform and applicable to a variety of tissue specimens.