BioSample

This library has been used successfully to clone a number of full-length cDNAs ranging in size from 1.4 to 4.5 kb. There are less than 0.5% clones with multiple inserts. Since each cDNA has an EcoRI site at its 5' end and an XhoI site at the 3' end, these clones can be easily identified. One should be suspicious of any clone which gives 3 or more bands in an EcoRI-XhoI double digest AND has an internal XhoI site. We usually do not further characterize any such clones unless the cDNA is known to give multiple bands in an EcoRI-XhoI digest. Microplate status: 500,000 unamplified cDNAs were mass excised (pBluescript SK-) in Xl1-Blue using ExAssist phage. The resulting single-stranded phagemids were used to infect Top10F'. Clones were picked into freezing medium (per liter 15 g tryptone, 10g yeast extract, 5g NaCl, 36 mM K2HPO4, 13.2 mM KH2PO4, 1.7 mM Na-citrate, 0.4 mM MgSO4*7 H2O, 6.8 mM (NH4)2SO4, 4% w/v glycerol) and grown for 24 hours. Original library contruction by Bruce Blumberg (Blumberg et al., 1991 Science 253, 194-196; Hawlet et al., 1995, Genes Dev. 9, 2923-2935). Note: This is a Xenopus Gene Collection (XGC) library