SAMN02230269
External Id
LIBEST_028248
External Id
University of New Hampshire
INSDC center name
2013-07-10T23:05:03.110Z
INSDC first public
2015-05-20T15:03:26.107Z
INSDC last update
live
INSDC status
Generic
NCBI submission model
Generic.1.0
NCBI submission package
smooth muscle cells
http://purl.obolibrary.org/obo/CL_0000192
cell type
The aortas from one-day old White Carneau (WC) pigeons were used to prepare primary smooth muscle cell cultures. Genetic expression was compared between Rosiglitazone-treated and control cells. Total RNA was extracted (Trizol, Invitrogen), purified, and converted to double-stranded cDNA using the BD SMART PCR cDNA Synthesis Kit (Clontech). Full-length cDNA from each treatment was cut with DpnII (5'GATC) in order to prepare amplicons for comparison by Representational Difference Analysis (RDA). cDNA fragments were subjected to three rounds of PCR-coupled subtractive hybridization and the final difference products were cloned into the BamHI restriction site of the pBluescript SK+ phagemid vector and transformed into XL1 MRF- chemically competent host cells (Stratagene). Transcripts were purified from the host cells by Rolling Circle Amplification (GE Healthcare) and sequenced by the DNAcore facility at the Hubbard Center for Genome Studies, University of New Hampshire. Sequences represent genes that were differentially expressed between the treatments. The WC_ROSI library contains transcripts that were upregulated in the Rosiglitazone-treated cells compared to the control cells. The corresponding library (WC_CONT) contains transcripts that were upregulated in the control cells compared to the Rosiglitazone-treated cells.
description
XL1 MRF-
lab-host
Columba livia
http://purl.obolibrary.org/obo/NCBITaxon_8932
organism
BamHI
re_1
mixed
http://www.ebi.ac.uk/efo/EFO_0001271
sex
White Carneau
strain
primary cell culture
http://purl.obolibrary.org/obo/OBI_0001910
tissue-type
WC_ROSI
title
pBluescript SK+
vector