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
CX675095CX675095EST
CX675096CX675096EST
CX675097CX675097EST
CX675098CX675098EST
CX675099CX675099EST
CX675100CX675100EST
CX675101CX675101EST
CX675102CX675102EST
CX675103CX675103EST
CX675104CX675104EST
CX675105CX675105EST
CX675106CX675106EST
CX675107CX675107EST
CX675108CX675108EST
CX675109CX675109EST
CX675110CX675110EST
CX675111CX675111EST
CX675112CX675112EST
CX675113CX675113EST
CX675114CX675114EST
CX675115CX675115EST
CX675116CX675116EST
CX675117CX675117EST
CX675118CX675118EST
CX675119CX675119EST

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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