Worm Breeder's Gazette 11(2): 17
These abstracts should not be cited in bibliographies. Material contained herein should be treated as personal communication and should be cited as such only with the consent of the author.
It is worth to obtain many kinds of cDNA molecules, regardless of identified or unidentified genes, in the form of independent clones. One of the useful approach for such a purpose is to construct a cDNA library which represents every gene uniformly, which makes it possible to analyze the clones by random selection. For this purpose,we have applied the genomic DNA hybridization to change the genetic population of cDNA library, for more even representation of the genes. First, we have made a vector-primed cDNA library from C. elegans polyA+ RNA fraction, and converted it to a single stranded form by helper phage infection (Figure). 7mg of the ssDNA was hybridized with 0.14mg of heat denatured C. elegans genomic DNA immobilized on a Nylon membrane filter. After extensive washing of the filter, the remaining ssDNA molecules were eluted with alkali. About 1 g of the ssDNA was recovered. The eluate was neutralized and used directly to transform E. coli host cells, which gave 5x10+E4 transformants. These transformants were found to be almost excluded from the empty clones, namely vector molecule itself with no cDNA, which shared nearly 80% in the original library. we are going to evaluate the new library doing differential hybridization of each clone. [See Figure 1]