BioSample

Library was prepared from 50 ug of total RNA by oligo-dT priming and AMV reverse transcriptase. After addition of EcoRI linkers and EcoRI-XhoI digestion, the cDNA was size selected by chromatography on Sepharose CL-4B columns and fractions containing cDNAs larger than 500 bp were ligated into EcoRI-XhoI-digested lambda ZAPII (UniZAP-XR) and packaged in vitro. Average insert size is 1.4 kb. The original library contained 6 x 106 recombinants, of which 3 x 106 were amplified and stored at -70 C in SM buffer containing 7% DMSO. 3 x 106 pfu were mass excised and the resulting phagemids used to infect Top10F. References: Science 253, 196-196 and Methods in Molecular Biology 97, 555-574. Additional sequences from this library have been deposited under the name Xenopus laevis dorsal blastopore lip. Library constructed by Bruce Blumberg (University of California, Irvine, Department of Developmental and Cell Biology).