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BioSample

National Center for Biotechnology Information

Organism: Xenopus tropicalis (tropical clawed frog)
cellular organisms; Eukaryota; Opisthokonta; Metazoa; Eumetazoa; Bilateria; Deuterostomia; Chordata; Craniata; Vertebrata; Gnathostomata; Teleostomi; Euteleostomi; Sarcopterygii; Dipnotetrapodomorpha; Tetrapoda; Amphibia; Batrachia; Anura; Pipoidea; Pipidae; Xenopodinae; Xenopus; Silurana

Attributes

tissue	whole embryo, pool of stages 10, 20 and 30
vector	pRKW2
v_type	phagemid (ampicillin resistant)
re_1	BamHI
re_2	XhoI

Description: 10 ug of polyA+ RNA was isolated from a mixture of embryos at stage 10, 20 and 30 and primed by oligo-dT primer: 5'-GAGAGAGAGAAGGATCC(T)16VN-3' (where V=G,A,C). 5-methyl-dCTP was used instead of dCTP in the first-strand synthesis in order to get hemimethylated cDNA. After full-length enrichment, oligo-dG tailing and normalization against itself, second-strand synthesis was carried out by priming with 5'-GAGAGAGAGACTCGAGTTAATTAAT(C)13-3'. dsDNA was digested with XhoI/BamHI and directionally cloned into the pRKW2 vector. Average insert size is 1.5 kb. Library constructed using the Carninci protocol (Genome Research 2000) by Drs. W. Wu and C. Niehrs (DKFZ, Heidelberg, Germany).