Parent Washington Navel Orange Callus cDNA Library UCRCS08-2

Overview
Library NameParent Washington Navel Orange Callus cDNA Library UCRCS08-2
Unique NameParent Washington Navel Orange Callus cDNA Library UCRCS08-2
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, Federici, Wanamaker, Lyon, Ye, Jang, Collin, Kacar, Ikeda, Quinitio). Sequences that survived all removal steps were submitted to GenBank.
Features
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Feature NameUnique NameType
CX077931CX077931EST
CX077930CX077930EST
CX077929CX077929EST
CX077928CX077928EST
CX077927CX077927EST
CX077926CX077926EST
CX077925CX077925EST
CX077924CX077924EST
CX077923CX077923EST
CX077922CX077922EST
CX077921CX077921EST
CX077920CX077920EST
CX077919CX077919EST
CX077918CX077918EST
CX077917CX077917EST
CX077916CX077916EST
CX077913CX077913EST
CX077912CX077912EST
CX077911CX077911EST
CX077910CX077910EST
CX077909CX077909EST
CX077908CX077908EST
CX077907CX077907EST
CX077906CX077906EST
CX077905CX077905EST

Pages

Properties
Property NameValue
Callustissue type
Genbank library cultivarWashington 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, Federici, Wanamaker, Lyon, Ye, Jang, Collin, Kacar, Ikeda, Quinitio). Sequences that survived all removal steps were submitted to GenBank.
Genbank library tissue typeCallus