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Nucleic Acids Research, 1992, Vol. 20, No. 14 3679-3684
© 1992

MOLECULAR BIOLOGY

Phenol-treatment and a homologous pairing-assay

Naoto Arai1,2, Katsumi Kawasaki1, Masa-aki Iwabuchi1 and Takehiko Shibata1,*

1Laboratory of Microbiology, The Insitute of Physical and Chemical Research(RIKEN) Wako-shi, Saitama 315–01 2The Graduate School of Science and Engineering, Saitama University Urawa-shi, Saitama 338, Japan

*To whom correspondence should be addressed

Received March 23, 1992. Accepted May 29, 1992.

Homologous pairing is a key step in homologous genetic recombination. In the early stage of trials for the Identification of homologous pairing-promoting proteins from a fission yeast, Schlzosaccharomyces pombe, we treated DNA products with phenol in the presence of a salt for the removal of tightly bound proteins from DNA before the assay, but we found that this treatment caused very efficient proteinindependent double-strand formation from complementary single-stranded DNAs. Using an assay including the phenol treatment, we detected another species of apparent homologous pairing-promoting proteins in the nuclei, in addition to a homologous pairing-promoting protein consisting of three components which we reported previously. However, studies involving the use of an assay without the phenol-treatments revealed that the second one was not really a homologous pairing-protein. Thus, the protein-independent double-strand formation by phenol-treatment in the presence of a salt could cause the erroneous identification of homologous pairingpromoting proteins.


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