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Nucleic Acids Research, 1994, Vol. 22, No. 6 987-992
© 1994

METHODS

Construction of a normalized cDNA library by introduction of a semi-solid mRNA-cDNA hybridization system

Yasnory F. Sasaki, Dai Ayusawa and Michio Oishi*

Institute of Molecular and Cellular Biosciences, The University of Tokyo Yayoi 1-1-1, Bunkyo-ku, Tokyo 113, Japan

* To whom correspondence should be addressed.

Received December 23, 1993. Revised February 16, 1994. Accepted February 16, 1994.

We report a novel procedure to construct a normalized (equalized) cDNA library. By introduction of the highly efficient self-hybrldlzatlon system between a whole mRNA population and their corresponding cDNA Immobilized on latex beads, which involves relatively simple manipulations, we were able to generate an mRNA population in which the copy number of abundant species was reduced while that of rare species was enriched. In a typical experiment, after several cycles of self-hybrldlzatlon on the beads, the ratio of the most to the least abundant marker mRNA species dropped by a factor of 300 (from 10,000 to 30) while the complexity and length of mRNAs in the population remained unchanged. The procedure should provide a potent tool for the expression cloning of cDNA and also facilitate the construction of whole cDNA catalogues from specific tissues (or cell types) from higher organisms.


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