Dataset EG_cDNA cDNA library

General Information
Name	EG_cDNA	Species	D. melanogaster
Dataset type	cDNA library	FlyBase ID	FBlc0000045
Source & Content
Consists of	cDNA clones in plasmid vector (Shigenobu et al., 2006)
Created by	Shuji Shigenobu, Okazaki National Research Institutes
Available from
Strain
Stage & tissue
Stage Tissue/Position (including subcellular localization) Reference embryonic stage 13 -- 16 gonad (Shigenobu et al., 2006)
Cell Line
Recent Updates
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FB2013_03
FB2013_02
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Description & Members
Description	Cloned cDNAs prepared from polyadenylated RNA isolated from embryonic gonads. (Shigenobu et al., 2006)
Parent collections
Component collection(s)
Number in collection
Comment on number in collection
Members
Experimental protocol
Vector	pDNR-LIB (Shigenobu et al., 2006) pGEMT-Easy (Shigenobu et al., 2006)
Sample preparation	Fly gonads were isolated from embryos carrying EGFP-vasa transgene that express GFP only in poll cells by using fluorescence-activated cell sorting (FACS). (Shigenobu et al., 2006)
Collection preparation	Total RNA was extracted from the isolated gonads at embryonic stages 13-16 by using RNeasy Mini kit (QIAGEN). Poly(A)+ RNA was purified Oligotex dT30 (TaKaRa) from the total RNA. cDNA was synthesized and amplified by Long-Distance PCR using SMART technology (Clontech) and either (1) cloned into pGEM-T Easy vector (Promega) cDNA inserts are flanked by linker sequences 5'-AAGCAGTGGTATCAACGCAGAGTGGCCATTACGGCCGGG-3' (5' linker: SMART IV Oligonucleotide, Clontech) and 5'-NBAAAAAAAAAAAAAAAAAAAAAAAAAAAAAACATGTCGGCCGCCTCGGCCTCTAGAAT-3' (3' linker: CDSIII/3' PCR Primer, Clontech) or (2) directionally cloned into pDNR-LIB vector (Clontech) cDNA inserts are flanked by linker sequences 5'-GGCCATTACGGCCGGG-3' (5' linker: SMART IV Oligonucleotide, Clontech) and 5'-polyA-CATGTCGGCCGCCTCGGCC-3' (3' linker: CDSIII/3' PCR Primer, Clontech). (Shigenobu et al., 2006)
Mode of assay
Assay platform
Data analysis
Additional data
	More information is available under: UniGene: D. melanogaster embryonic gonads cDNA, pDNR-LIB UniGene: D. melanogaster embryonic gonads cDNA, pGEM-T Easy
Associated files
Additional sites
Synonyms & Secondary IDs
Reported As
Symbol Synonym	EG (Shigenobu et al., 2006) EG_cDNA
Secondary FlyBase IDs

References ( 1 )
Research paper	Shigenobu et al., 2006, Proc. Natl. Acad. Sci. U.S.A. 103(37): 13728--13733 Molecular characterization of embryonic gonads by gene expression profiling in Drosophila melanogaster. [FBrf0193273]