|
Status |
Public on Sep 08, 2009 |
Title |
LM6, biological rep1 |
Sample type |
RNA |
|
|
Source name |
Human lipoma-derived mesenchymal stem cells
|
Organism |
Homo sapiens |
Characteristics |
Gender: female
|
Growth protocol |
LM6 cells and AM3 cells were cultured in the K-NAC medium that is a modified MCDB 153 medium (Keratinocyte-SFM, GIBCO-Invitrogen) supplemented with N-acetyl-L-cysteine (NAC; Sigma A8199) (2 mM) and L-ascorbic acid 2-phosphate (Asc 2P; sigma A8960) (0.2 mM).
|
Extracted molecule |
total RNA |
Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions. Total RNAs extracted using TRIZOL reagent were quantified spectrophotometrically. The cDNA synthesis was carried out by using SUPERSCRIPTTM one-step RT-PCR kit (Invitrogen). For the reverse transcription step, the whole 5 ul of the resuspended RNAs were incubated for 60 min at 42oC, then 15 min at 72oC in 50 ul of reaction mixture containing 25 ul of 2x Reaction Mix (Invitrogen) and 1 ul of RT/PLANTINUM Taq Mix (Invitrogen). 28.5 ul of the cDNAs present in the 50 ul RT reaction mixture purified by Microarray Target Purification Kit (Roche Applied Science, Indianapolis, ID, USA, http://www.roche-applies-science.com) were used as templates to amplify cDNA by Microarray Target Amplification Kit. The amplified cDNAs were purified by Microarray Target Purification Kit, and complementary RNAs (cRNAs) were synthesized from 200 ng cDNA with Microarray RNA Target Synthesis Kit (Roche Applied Science).
|
Label |
biotin
|
Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 5 micrograms purified cRNA (Expression Analysis Technical Manual, 2004, Affymetrix).
|
|
|
Hybridization protocol |
Following fragmentation, 10 micrograms of cRNA were hybridized for 16 hr at 45oC on Affymetrix Human Genome U133 plus 2.0 GeneChip. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
Scan protocol |
GeneChips from the hybridization experiments were read by the Affymetrix GeneChip scanner 3000.
|
Description |
The mRNA profiles from LM6 cells and AM1cells were quantitatively determined.
|
Data processing |
The data were processed by GeneSpring software version 7.2 GC-RMA preprocessor to use on the CEL files
|
|
|
Submission date |
Sep 18, 2008 |
Last update date |
Aug 15, 2013 |
Contact name |
Sung-Liang Yu |
E-mail(s) |
slyu@ntu.edu.tw
|
Phone |
886-2-23958341
|
Organization name |
National Taiwan University
|
Department |
Clinical Laboratory Sciences and Medical Biotechnology
|
Lab |
Microarray Core Facility
|
Street address |
Jen Ai Road Section1
|
City |
Taipei |
ZIP/Postal code |
100 |
Country |
Taiwan |
|
|
Platform ID |
GPL570 |
Series (1) |
GSE12843 |
Comparative profiling of mRNA expression in human mesenchymal stem cells derived from adult adipose and lipoma tissues |
|
Relations |
Reanalyzed by |
GSE20125 |
Reanalyzed by |
GSE49910 |