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A lambda cDNA library from the human 293 cell line, which has high levels of telomerase activity, was partitioned into 25 pools containing ∼200,000 plaques each. These were screened by PCR with primers LT5 (CGGAAGAGTGTCTGGAGCAA) and LT6 (GGATGAAGCGGAGTCTGGA) to the 5′ region of the #712562 clone insert. Six subpools of one positive primary pool were further screened by PCR. One positive subpool was then screened by plaque hybridization with a probe from the 5′ region of clone #712562. One phage was positively identified and the ∼4 kbp insert from this clone was excised and subcloned into the pBluescript II SK+ vector (Stratagene) as an Eco RI fragment.
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We thank R. Adams, B. Lastelic, L. Tonkin, and F. Wu for expert technical assistance; C. Chapon, J. P. Cooper, R. Gutell, E. Jabri, and J. Sperger for discussions; R. Allshire and J. A. Wise for plasmids and yeast strains; C. Mattison and the L. Pillus lab for help with microscopy; and A. Sirimarco for manuscript preparation. An S. pombe cDNA library was provided by C. J. Norbury and B. Edgar. Supported by NIH grant GM28039 (T.R.C).
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