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cDNA was prepared from 2ug of poly A+ RNA. EcoRI-XhoI cut cDNA was then ligated into UniZap-XR (Stratagene) with EcoRI at the 5' end and XhoI at the 3' end. SS-library phagemids were prepared by mass excision from the original library and normalized by hybridization to biotinylated driver (prepared from the same library by PCR) to Cot-omega of 11. After removal of hybrids and excess driver by streptavidin sepharose chromatography, the ss-phagemids were made double stranded and electroporated into Top-10 F'. Original library contruction by Bruce Blumberg (Blumberg et al., 1991 Science 253, 194-196; Hawlet et al., 1995, Genes Dev. 9, 2923-2935). Normalized by Jihwan Song (Song, Cho and Blumberg, unpublished). Note: This is a Xenopus Gene Collection (XGC) library.
Nucleotide
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