Detection of point mutations by non-isotopic single strand conformation polymorphism

Brazilian Journal of Medical and Biological Research

N.a. Pinheiro²  R.p. Moura¹  E. Monteiro¹  L.l. Villa¹ 

[1] ,Universidade de São Paulo

关键词: p53 gene;    human papillomavirus;    PCR;    SSCP;    cancer;    silver staining;   

DOI  :  10.1590/S0100-879X1999000100008

来源: SciELO

【 摘 要 】

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.

【 授权许可】

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