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GEO help: Mouse over screen elements for information. |
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| Status |
Public on Dec 31, 2016 |
| Title |
wa_6h miRNA array |
| Sample type |
RNA |
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| Source name |
3T3-L1 cells
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| Organism |
Mus musculus |
| Characteristics |
day of differentiation: day 0, 6h
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| Treatment protocol |
NA
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| Growth protocol |
For C3H10T1/2: Growth medium: DMEM (high glucose), 10% Fetalclone III serum, PenStrep; Induction medium (d0-d2): DMEM (high glucose), 10% Fetalclone III serum, PenStrep, 5uM DEX, 0.5mM IBMX, 0.12ug/ml Insulin, 1uM Rosi, 1nM T3; Differentiation medium (d2-d7): DMEM (high glucose), 10% Fetalclone III serum, PenStrep, 0.12ug/ml Insulin, 1uM Rosi, 1nM T3. Before differentiation cells were seeded at 70% confluency and growth medium was supplemented with 8.3nM human recombinant BMP7 for 3 days.For 3T3-L1: Growth medium: DMEM (high glucose), 10% BCS, PenStrep; Induction medium (d0-d2): DMEM (high glucose), 10% FBS, PenStrep, 5uM DEX, 0.5mM IBMX, 0.12ug/ml Insulin; Differentiation medium (d2-d7): DMEM (high glucose), 10% FBS, PenStrep, 0.12ug/ml Insulin. Before differentiation cells were seeded at 70% confluency (d-4).
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| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extrated using Trizol reagent following the manufactorer's instructions and treated with DnaseI.
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| Label |
Hy3
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| Label protocol |
For C3H10T1/2: 550 ng total RNA from samples was labeled with Hy3™ fluorescent label, respectively, using miRCURY LNA™ microRNA Hi-Power Labeling Kit, Hy3™ (Exiqon, Denmark) following the procedure described by the manufacturer. The Hy3™-labeled samples were hybridized to the miRCURY LNA™ microRNA Array 7th Gen (Exiqon, Denmark), which contains capture probes targeting all microRNAs for human, mouse or rat registered in the miRBASE 18.0. For 3T3-L1: for labelling the miRCURY LNA™ microRNA Power Labeling Kit (Hy3) was used follwoing the manufatorer's instructions and hybridized to the miRCURY LNA™ microRNA Array 4th Gen (Exiqon, Denmark).
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| Hybridization protocol |
The hybridization was performed according to the miRCURY LNA™ microRNA Array Instruction manual using a Tecan HS4800™ hybridization station (Tecan, Austria). For C3H10T1/2: Array product number 208500, Array version 7th Gen, Array batch number 35001, miRBase version 18. For 3T3-L1 Array product number 208200, Array version 4th Gen, Array batch number 31028.
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| Scan protocol |
For C3H10T1/2: After hybridization the microarrayslides were scanned and stored in an ozone free environment (ozone level below 2.0 ppb) order to prevent potential bleaching of the fluorescent dyes. The miRCURY LNA™ microRNA Array slides were scanned using the Agilent G2565BA Microarray Scanner System (Agilent Technologies, Inc., USA) and the image analysis was carried out using the ImaGene® 9 (miRCURY LNA™ microRNA Array Analysis Software, Exiqon, Denmark). For 3T3-L1: microarrayslides were scanned using GenePix 4000P.
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| Description |
Growth medium: DMEM (high glucose), 10% BCS, PenStrep; Induction medium (d0-d2): DMEM (high glucose), 10% FBS, PenStrep, 5uM DEX, 0.5mM IBMX, 0.12ug/ml Insulin; Differentiation medium (d2-d7): DMEM (high glucose), 10% FBS, PenStrep, 0.12ug/ml Insulin. Before differentiation cells were seeded at 70% confluency (d-4).
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| Data processing |
The microRNA array signals were quantile normalized and then log2 transformation
R was used for quantile normalization
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| Submission date |
Dec 04, 2015 |
| Last update date |
Dec 31, 2016 |
| Contact name |
Wei Xie |
| E-mail(s) |
[email protected]
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| Organization name |
Tsinghua University
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| Street address |
zhongguancun beidajie
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| City |
Beijing |
| ZIP/Postal code |
100086 |
| Country |
China |
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| Platform ID |
GPL21206 |
| Series (2) |
| GSE75697 |
miRNA expression during differentiation of white and brown adipocytes |
| GSE75698 |
Comparative transcriptomic and epigenomic analyses reveal new regulators of murine brown adipogenesis |
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| Supplementary file |
Size |
Download |
File type/resource |
| GSM1964306_wd0_6_array.txt.gz |
409.9 Kb |
(ftp)(http) |
TXT |
| Processed data are available on Series record |
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