BioSample

cDNA was prepared from 2ug of poly A+ RNA (equal parts from stage 10.5 and stage 11.5 gastrulae). 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 (Cho et al. 1991 Cell 67, 1111-1120). Normalized by Jihwan Song (Song, Cho and Blumberg, unpublished). Note: This is a Xenopus Gene Collection (XGC) library.