|
| Status |
Public on Jun 30, 2023 |
| Title |
HM cell line |
| Sample type |
RNA |
| |
|
| Source name |
The human melanocytes HM
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: melanocytes
cell line: HM
|
| Treatment protocol |
The cells were incubated with Dulbecco's modified Eagle's medium containing 10% fetal bovine serum in a 5% CO2 incubator at 37˚C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the cells using the RNA extraction kit according to the manufacturer's instructions and examined by 1% agarose gel electrophoresis.The purity and concentration of total RNA samples were determined with NanoDrop ND-1000.
|
| Label |
Cy3
|
| Label protocol |
Sample labeling and array hybridization were performed according to the manufacturer’s protocol (Arraystar Inc.). Briefly,circRNA weretreated with Rnase R (Epicentre, Inc.) to remove linear RNAs. Then, each sample was amplified and transcribed into fluorescent cRNA utilizing a random priming method (Arraystar Super 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 each 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 × Hybridization buffer was added to dilute the labeled cRNA. 50 μl of hybridization solution was dispensed into the gasket slide and assembled to the circRNA 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 using the Agilent Scanner G2505C.
|
| Data processing |
Agilent Feature Extraction software (version 11.0.1.1) was used to analyze acquired array images. Quantile normalization and subsequent data processing were performed using the R software package.Differentially expressed circRNAs between groups was screened by fold change and P-value.
|
| |
|
| Submission date |
Jun 22, 2023 |
| Last update date |
Jun 30, 2023 |
| Contact name |
Qi Wang |
| E-mail(s) |
wangrock2005@sina.com
|
| Phone |
13077332109
|
| Organization name |
The Third Xiangya Hospital of Central South University
|
| Street address |
Tongzipo Road
|
| City |
Chang Sha |
| State/province |
Other (Non U.S.) |
| ZIP/Postal code |
410013 |
| Country |
China |
| |
|
| Platform ID |
GPL19978 |
| Series (1) |
| GSE235561 |
Analysis of circRNAs expression in melanocytes and melanoma cells |
|
