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Sample GSM1388224 Query DataSets for GSM1388224
Status Public on May 28, 2014
Title miRNA_Control ASM cells_rep3
Sample type RNA
Source name ASM, 24hr
Organism Homo sapiens
Characteristics tissue: Airway Smooth Muscle
Growth protocol Confluent cells were growth-arrested by FCS deprivation for 24 h before being treated with dexamamethasone for 60 min, and then stimulated with FCS for a further 24 h.
Extracted molecule total RNA
Extraction protocol Total RNA was extracted using the mirVana™miRNA isolation kit (Ambion Europe). RNA was eluted in 50 μl RNase-free water (Promega UK, Southampton, UK) and stored at -80 °C. RNA content and purity was measured using a BioTek PowerWave XS (SSi Robotics, Tustin, CA, U.S.A.) spectrophotometer.
Label Cy3
Label protocol miRNA samples were labeled using cy3
Hybridization protocol miRNA-enriched RNA samples (100 ng) used in the microRNA microarrays were labeled with the Labeling Spike-In solution using Agilent’s miRNA Complete Labeling and Hyb Kit and then the samples were dephosphorylated by incubation at 37 oC on a heat block for 30 min. DMSO was added to the dephosphorylated RNA for denaturation for 10 min, followed by the assembly of the Labeling Reaction, where samples were incubated with Cyanine 3-pCp and allowed to ligate for 2 h at 16 oC. The labeled RNA was purified using Micro Bio-Spin 6 columns (Bio-Rad) and the desalted samples were dried up in a vacuum concentrator at 50 oC for 1 h to remove DMSO. Samples were resuspended in Hybridization Mix using Hyb Spike-In solution and Hybridization Buffer and were loaded onto Agilent Human miRNA microarrays and hybridized at 55 oC for 20 h at 20 rpm using Agilent’s Hybridization oven and SureHyb chamber. The microarrays were then disassembled and washed in GE Wash Buffer 1 (Agilent) for 60 sec at RT, followed by GE Wash Buffer 2 for 60 sec at 37 oC. The microarrays were scanned with the Agilent Microarray Scanner G2565BA using the profile for miRNA microarrays (AgilentHD_miRNA) at 2 μm resolution, dye channel Green, Scan Area 61 x 21.6 mm.
Scan protocol Scanned on an Agilent Sureprint G3 scanner.
Description Gene expression after 24h
Data processing Agilent Feature Extraction Software
Submission date May 16, 2014
Last update date May 28, 2014
Contact name Mark Perry
Organization name NHLI, Imperial College London
Department Airway Disease
Street address Dovehouse Street
City London
ZIP/Postal code SW36LY
Country United Kingdom
Platform ID GPL15018
Series (2)
GSE57765 Gene expression analysis of stimulated primary ASM cells [microRNA]
GSE57766 Gene expression analysis of stimulated primary ASM cells

Data table header descriptions
VALUE Normalized signal

Data table
1 1.15E+02
2 -1.63E+01
3 -1.75E+01
4 -1.84E+01
5 -2.07E+01
6 -1.90E+01
7 -1.82E+01
8 -1.71E+01
9 -1.57E+01
10 -1.68E+01
11 3.98E+00
12 -2.22E+00
13 -1.86E+00
14 -1.38E+00
16 1.47E+00
17 1.34E+01
19 -2.28E+00
20 1.52E+01
21 1.93E+00
22 -2.75E+00

Total number of rows: 56044

Table truncated, full table size 833 Kbytes.

Supplementary file Size Download File type/resource
GSM1388224_US91903684_253118110674_S02_miRNA_105_Dec08_1_4.txt.gz 8.1 Mb (ftp)(http) TXT
Processed data included within Sample table

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