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NIA Mouse E13.5 VMB Dopamine cell cDNA Library

BioSample: SAMN00160791; EST: LIBEST_006255
Mus musculus (house mouse)
cellular organisms; Eukaryota; Opisthokonta; Metazoa; Eumetazoa; Bilateria; Deuterostomia; Chordata; Craniata; Vertebrata; Gnathostomata; Teleostomi; Euteleostomi; Sarcopterygii; Dipnotetrapodomorpha; Tetrapoda; Amniota; Mammalia; Theria; Eutheria; Boreoeutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; Murinae; Mus; Mus
strainTH-beta-gal transgenic mouse
development stage13.5dpc
lab hostDH10B
vectorpSPORT1 (Gibco/BRL Life Technology)
v_typeplasmid (ampicillin resistant)

Total RNAs were extracted from 3000 Dopamine cells (cell collected by Dr.Tanya Barrett). The double-stranded cDNA was synthesized by Gibco's kit with an Oligo(dT) primer [NotI primer-adapter from GibcoBRL] [5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 0.91ug of total RNA. The double-stranded cDNAs were treated with T4 DNA polymerase and purified by ethanol-precipitation. The cDNAs were ligated to Lone-linker LL-Sal4 (include Sal1 sequence). The cDNAs were purified by phenol/chloroform and separated from free linkers by Centricon 100. Then, cDNAs were amplified by long-range high fidelity PCR using Takara's Ex Taq polymerase. Then, the cDNAs were purified by phenol/chloroform and by Centricon 100. The cDNAs were digested with SalI and NotI enzymes. Then, the cDNAs were size selected by Gibco's Size Fractionation Column. The cDNAs were cloned into SalI/NotI site of pSPORT1 plasmid vector. The DH10B E. coli host was transformed with the ligation mixture by chemical method. The library was constructed by Yulan Piao and Minoru.S. H. Ko.

National Institute on Aging/National Institutes of Health, Dawood B. Dudekula; 2000-09-21

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