Washington Navel orange cold acclimated flavedo & albedo cDNA library

Vector: Lambda Uni-ZAP XR, excised phagemid; Site_1: EcoRI; Site_2: XhoI; Plants were grown in the field at University of California, Riverside Agricultural Operations since 1983. The scion was Washington Navel orange and the rootstock Carizzo Citrange. Tissue from mature fruit was harvested at mid-day in January 2002 during a cold spell, when pre-dawn temperatures were approximately -2 to 2 degree C. Approximately 2 cm median sections of the rind were excised in the field from several fruits, then wrapped in aluminum foil and frozen quickly in dry ice. Total RNA was extracted using a phenol extraction procedure described in J. Japanaese Soc. Hort. Sci. (1996) 64:809-814. Poly(A) RNA was purified, a cDNA library was made, and 1 million primary lambda cDNA clones were in vivo excised to give a population of pBluescript SK(-) phagemids. All steps to this point were performed in the TJ Close lab at the University of California, Riverside (Fenton). Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3530 at the Arizona Genomics Institute, University of Arizona (Collura, Feuerbacher, Kim, Kudrna, Wing, Yu). Chromatogram files were transmitted to UC Riverside (by Yu), then processed at UC Riverside (by Wanamaker) using the HarvEST pipeline (http://harvest.ucr.edu) to remove vector and cloning oligo sequences and various contaminants, and to trim to a high quality region. Sequences that retained a phred 17 region of at least 100 bases were deposited to GenBank.