|
| Status |
Public on Nov 28, 2024 |
| Title |
Circular RNA_siSMG6_CBP80IP_biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
HeLa
|
| Organism |
Homo sapiens |
| Characteristics |
gender: Female
treatment: siSMG6 + CBP80 IP
|
| Treatment protocol |
RNA immunoprecipitations were performed using either α-CBP80 or α-eIF4E antibodies.
|
| Growth protocol |
HeLa cells were grown with Dulbecco’s Modified Eagles Medium (Sigma-aldrich) supplemented with 10% (vol/vol) fetal bovine serum (Sigma-aldrich) and 1% (vol/vol) penicillin/streptomycin (Sigma-aldrich)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs associated with CBP80 or eIF4E were extracted using Trizol by manufacturer's recommendations
|
| Label |
Cy3
|
| Label protocol |
The enriched circular RNAs were amplified and transcribed into Cy3 labeled cRNA utilizing a random priming method (Arraystar Super RNA Labeling Kit; Arraystar). The labeled cRNAs were purified by RNeasy Mini Kit (QIAGEN)
|
| |
|
| 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. The hybridized arrays were washed, fixed and scanned using the Agilent Scanner G2505C.
|
| Scan protocol |
The signals were scanned using the Agilent Scanner G2505C
|
| Description |
Circular RNA_siSMG6_CBP80IP_biological rep2
|
| Data processing |
Quantification was performed by Feature Extraction Software (Agilent Technology, Palo Alto, CA), and analysis were performed using GeneSpringGX 7.3 (Agilent Technology, USA)
|
| |
|
| Submission date |
Aug 03, 2023 |
| Last update date |
Nov 28, 2024 |
| Contact name |
Sung Ho Boo |
| Organization name |
Korea Advanced Institute of Science and Technology
|
| Street address |
291, Daehak-ro
|
| City |
Daejeon |
| ZIP/Postal code |
KS015 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL21825 |
| Series (2) |
| GSE239958 |
Enrichment of CBP80-associated and eIF4E-associated circular RNAs in SMG6 depleted cells |
| GSE240092 |
Circular RNAs affect mRNA stability |
|
