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S9SNU601

Identifiers
BioSample: SAMN00170204; EST: LIBEST_010309
Organism
Homo sapiens (human)
cellular organisms; Eukaryota; Opisthokonta; Metazoa; Eumetazoa; Bilateria; Deuterostomia; Chordata; Craniata; Vertebrata; Gnathostomata; Teleostomi; Euteleostomi; Sarcopterygii; Dipnotetrapodomorpha; Tetrapoda; Amniota; Mammalia; Theria; Eutheria; Boreoeutheria; Euarchontoglires; Primates; Haplorrhini; Simiiformes; Catarrhini; Hominoidea; Hominidae; Homininae; Homo
Attributes
sexM
tissueAscites
cell typeEpithelial
cell lineSNU-601
lab hostTop10F'
tissueStomach
vectorpME18-FL3
v_typePlasmid
re_1XhoI
re_2XhoI
Description

The poly (A)+ RNA was dephosphorylated with bacterial alkaline phosphatase (BAP) and then decapped with tabacco acid pyrophosphatase (TAP). The decapped intact mRNA was ligated with DNA-RNA linker including SfiI site by treatment of T4 RNA ligase and the first strand cDNA was synthesized with Superscript II using SfiI oligo-dT primer. After first strand synthesis, RNA was degraded by NaOH treatment and cDNA was amplified by PCR reaction. The PCR products were digested with SfiI and cloned into DraIII- digested pME18S-FL3 vector. The obtained cDNA vectors were used for transformation of competent cells E. coli Top10F' by electroporation method. The cDNA libraries constructed by this method are full-length enriched cDNA library.

Submission
Korea Research Institute of Bioscience & Biotechnology, Kim YS; 2002-03-01
Accession:
SAMN00170204
ID:
170204

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