|
| Status |
Public on Sep 29, 2022 |
| Title |
DM1 |
| Sample type |
RNA |
| |
|
| Source name |
full thickness skin of 8-week old STZ induced SD male rat
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague-Dawley (SD)
tissue: full thickness skin
treatment: intraperitoneally (i.p.) injected with 60 mg/kg streptozotocin
|
| Treatment protocol |
4-week old Sprague-Dawley (SD) male rats were intraperitoneally (i.p.) injected with 60 mg/kg streptozotocin to induce diabetes and were housed for 4 weeks before being anesthetized, and full-thickness skin tissue was harvested for microarray analysis.
Four-week-old male Sprague Dawley (SD) rats (weighing 250-300 g) were purchased from the Laboratory Animal Center of Sun Yat-sen University. the animals were i.p. injected with 60 mg/kg streptozotocin (STZ) to induce diabetes and allowed to manifest hyperglycemia for 4 weeks before inflicting a cutaneous wound.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the cells using TRIzol reagent (AGbio, China) following the manufacturer’s instructions.RNA quantity and quality were measured by NanoDrop ND-1000. RNA integrity was assessed by standard denaturing agarose gel electrophoresis.
|
| Label |
Cy3
|
| Label protocol |
Sample labeling were performed according to the Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technology). Briefly, total RNA from each sample was linearly amplified and labeled with Cy3-UTP. The labeled cRNAs were purified by RNeasy Mini Kit (Qiagen). The concentration and specific activity of the labeled cRNAs (pmol Cy3/μg cRNA) were measured by NanoDrop ND-1000.
|
| |
|
| Hybridization protocol |
Array hybridization were performed according to the Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technology). 1 μg of each labeled cRNA was fragmented by adding 11 μl 10 × Blocking Agent and 2.2 μl of 25 × Fragmentation Buffer, then heated the mixture at 60°C for 30 min, finally 55 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 100 μl of hybridization solution was dispensed into the gasket slide and assembled to the gene expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven.
|
| Scan protocol |
The hybridized arrays were washed, fixed and scanned with using the Agilent DNA Microarray Scanner (part number G2505C).
|
| Description |
full thickness skin of 8-week old STZ induced SD male rat replicate1
|
| Data processing |
the scanned images were analysed using Agilent Feature Extraction software (v11.0.0.1).
Normalization of signal Raw signal intensities were normalized in quantile method by GeneSpring GX v11.5.1, and low intensity LncRNAs were filtered intensity using Agligent GeneSpring GX v12.1 software
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| |
|
| Submission date |
Sep 28, 2022 |
| Last update date |
Sep 29, 2022 |
| Contact name |
Meng Ren |
| E-mail(s) |
renmeng80@139.com
|
| Phone |
+862081332286
|
| Organization name |
Sun Yat-sen Memorial Hospital, Sun Yat-sen University
|
| Department |
endocrinology department
|
| Street address |
107 Yanjiang West Road
|
| City |
guangzhou |
| ZIP/Postal code |
510120 |
| Country |
China |
| |
|
| Platform ID |
GPL15690 |
| Series (1) |
| GSE214338 |
Development of LncRNAs expression signisure of STZ induced rats skin |
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