Providing means to an end

Eisenstein, Michael

doi:10.1038/nmeth0805-628

Classic Protocol
Published: August 2005

Providing means to an end

Michael Eisenstein

Nature Methods volume 2, page 628 (2005)Cite this article

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By the mid-1980s, the cloning era was entering full bloom. cDNA synthesis techniques were finding widespread use, and many researchers were taking advantage of the newly obtained ability to generate immense cDNA libraries for the identification of novel genes. “The field, the way it was in the eighties, was a lot of fun,” recalls Michael Frohman, at the State University of New York at Stony Brook. “You would clone a gene and figure out the sequence and do some in situ hybridization and see what the gene expression pattern was, and then the next part was really hard—you had to do something to figure out what the function was—so most people just gave up at that point and said, 'well, there are lots more genes to clone!'”

Of course, cloning wasn't always just fun and games—conventional cDNA cloning techniques often didn't make it all the way to the 5′ end of a given transcript, and many scientists found themselves frustrated in their efforts to obtain full-length cDNA sequences from long or scarce transcripts. Frohman encountered this as a postdoc in the lab of Gail Martin, at the University of California at San Francisco, where he was struggling as a molecular biology neophyte to clone the full-length engrailed gene. “It was a typical story for the time,” recalls Frohman. “Everybody was making cDNA libraries and screening for low-abundance genes; they were difficult to clone, you would maybe get a couple of hits after plating out twenty huge plates of phage, and frequently it wasn't full-length, and you'd have to start over. It was a frustrating thing, and people would spend six months to a year, to two years, unable to do anything else because they couldn't get a full-length gene product.”

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References

Saiki, R.K. et al. Science 230, 1350–1354 (1985).
Article CAS Google Scholar
Mullis, K. et al. Cold Spring Harb. Symp. Quant. Biol. 51, 263–273 (1986).
Article CAS Google Scholar
Frohman, M.A. et al. Proc. Natl. Acad. Sci. USA 85, 8998–9002 (1988).
Article CAS Google Scholar
Loh, E.Y. et al. Science 243, 217–220 (1989).
Article CAS Google Scholar
Ohara, O. et al. Proc. Natl. Acad. Sci. USA 86, 5673–5677 (1989).
Article CAS Google Scholar