Parent Washington Navel Orange Callus cDNA Library UCRCS08-3

Overview
Library NameParent Washington Navel Orange Callus cDNA Library UCRCS08-3
Unique NameParent Washington Navel Orange Callus cDNA Library UCRCS08-3
OrganismCitrus sinensis (Sweet orange)
Typecdna_library
Vector: Lambda Uni-ZAP XR, excised phagemid; Site_1: EcoRI; Site_2: XhoI; Parent Washington navel orange embryogenic callus was established from undeveloped ovules of ca. 10 mm diameter young fruits under open pollination on Murashige-Skoog medium at 25oC with 16 h light in a tissue culture room. Embryogenic callus, globular and heart stage embryoids were pooled in approximately equal portions in RNAlater (Ambion), then RNA was extracted using TRIZOL Reagent (Invitrogen). Poly(A) RNA was purified from 500 microgram of total RNA using Qiagen Oligotex. A primary cDNA library was produced using a lambda ZAP XR cDNA Synthesis Kit (Stratagene). These steps were performed by Xinrong Ye (Roose lab, UC Riverside). One million pfu from the primary library were mass excised to produce a phagemid population by Raymond Fenton (Close lab, UC Riverside). Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at DNA Landmarks (Landry, Hubert, Laforest, Landry, Ligonde). Chromatogram files were downloaded by FTP to UC Riverside (by Close), then processed at UC Riverside (by Wanamaker, Close lab) 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 assembled, then chimeras were removed following manual inspection of assemblies (Close, Roose, Wanamaker). Sequences that survived all removal steps were submitted to GenBank.
SNP Chip Base
Array NameParent Washington Navel Orange Callus cDNA Library UCRCS08-3
OrganismCitrus sinensis (Sweet orange)
Typecdna_library
Features
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Feature NameUnique NameType
CX675070CX675070EST
CX675071CX675071EST
CX675072CX675072EST
CX675073CX675073EST
CX675074CX675074EST
CX675075CX675075EST
CX675076CX675076EST
CX675077CX675077EST
CX675078CX675078EST
CX675079CX675079EST
CX675080CX675080EST
CX675081CX675081EST
CX675082CX675082EST
CX675083CX675083EST
CX675084CX675084EST
CX675085CX675085EST
CX675086CX675086EST
CX675087CX675087EST
CX675088CX675088EST
CX675089CX675089EST
CX675090CX675090EST
CX675091CX675091EST
CX675092CX675092EST
CX675093CX675093EST
CX675094CX675094EST

Pages

Properties
Property NameValue
Callustissue type
Genbank library cultivarParent Washington Navel
Genbank library dev stageEmbryogenic and embryoid
Genbank library noteVector: Lambda Uni-ZAP XR, excised phagemid; Site_1: EcoRI; Site_2: XhoI; Parent Washington navel orange embryogenic callus was established from undeveloped ovules of ca. 10 mm diameter young fruits under open pollination on Murashige-Skoog medium at 25oC with 16 h light in a tissue culture room. Embryogenic callus, globular and heart stage embryoids were pooled in approximately equal portions in RNAlater (Ambion), then RNA was extracted using TRIZOL Reagent (Invitrogen). Poly(A) RNA was purified from 500 microgram of total RNA using Qiagen Oligotex. A primary cDNA library was produced using a lambda ZAP XR cDNA Synthesis Kit (Stratagene). These steps were performed by Xinrong Ye (Roose lab, UC Riverside). One million pfu from the primary library were mass excised to produce a phagemid population by Raymond Fenton (Close lab, UC Riverside). Phagemids were plated, plasmid DNA purified, cDNA clones archived, and DNA sequences determined bi-directionally using an ABI3730 at DNA Landmarks (Landry, Hubert, Laforest, Landry, Ligonde). Chromatogram files were downloaded by FTP to UC Riverside (by Close), then processed at UC Riverside (by Wanamaker, Close lab) 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 assembled, then chimeras were removed following manual inspection of assemblies (Close, Roose, Wanamaker). Sequences that survived all removal steps were submitted to GenBank.
Genbank library tissue typeCallus