|
| Status |
Public on May 28, 2021 |
| Title |
control group lncRNA 1 |
| Sample type |
RNA |
| |
|
| Source name |
cell line C2C12
|
| Organism |
Mus musculus |
| Characteristics |
differentiation: myotube
cell line: C2C12
treatment: none
|
| Treatment protocol |
C2C12 was cultured in DMEM high glucose supplemented with 10% fetal bovine serum, 100 U/mL of penicillin, and 100 μg/mL of streptomycin in 5% CO2 at 37°C. When the cells reached 80–90% confluence, they were differentiated by incubation in DMEM containing 2% horse serum. CoCl2 (200 uM) were added to cells for 48 h prior to cell collection.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cells using TRIZOL reagent (Life Technologies, Grand Island, NY, USA) in accordance with the manufacturer’s protocol. RNA quantity and quality were measured by NanoDrop ND-1000 (Thermo Fisher Scientific, Waltham, MA, USA). RNA integrity was assessed by standard denaturing agarose gel electrophoresis.
|
| Label |
Cy3
|
| Label protocol |
mRNA was purified from total RNA after removal of rRNA (mRNA-ONLY™ Eukaryotic mRNA Isolation Kit, Epicentre). Then, each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 3’ bias utilizing a random priming method (Arraystar Flash RNA Labeling Kit, Arraystar). 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 |
1 μg of Cy3-labeled cRNA was fragmented by adding 5 μl 10 × Blocking Agent and 1 μl of 25 × Fragmentation Buffer, then heated the mixture at 60 °C for 30 min, finally 25 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 50 μl of hybridization solution was dispensed into the gasket slide and assembled to the LncRNA 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).
|
| Data processing |
The scanned images were analyzed with Agilent feature extraction software (version 11.0.1.1).
|
| |
|
| Submission date |
May 30, 2018 |
| Last update date |
May 28, 2021 |
| Contact name |
Rui Chen |
| E-mail(s) |
rui.c.med@163.com
|
| Organization name |
Guangdong Second Provincial General Hospital
|
| Department |
Guangdong Traditional Medical and Sports Injury Rehabilitation Research Institute
|
| Street address |
466 Xin Gang Zhong Road
|
| City |
Guangzhou |
| ZIP/Postal code |
510317 |
| Country |
China |
| |
|
| Platform ID |
GPL19286 |
| Series (2) |
| GSE115089 |
CoCl2 induced cytotoxicity is associated with dysregulated lncRNAs expression |
| GSE115090 |
CoCl2 induced cytotoxicity is associated with dysregulated circRNAs and lncRNAs expression |
|
