|
| Status |
Public on Mar 02, 2021 |
| Title |
Bcl-xAS-transfected 786-O cells_replicate 2_Cy3 vs. Control empty vector-transfected 786-O cells_replicate 2_Cy5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Control empty vector-transfected 786-O cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: 786-O
original tissue: clear cell renal cell cancer (CCRCC)
transfection: control
|
| Treatment protocol |
Cells were transfected with 6 μg of Bcl-xAS expression construct or with 6 μg of empty vector (pCEP) as control using 3:1 FuGENE HD reagent (Promega). At 24 h after transfection, total RNA was extracted.
|
| Growth protocol |
786-O cells were cultivated in 147.8 cm2 cell culture plates with Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS. To perform four biological replicates, we cultivated four plates in parallel.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol reagent following the manufacturer's instructions. Total RNA was DNase-I-treated and purified using the Illustra RNAspin Mini kit (GE Healthcare) following the manufacturer's instructions.
|
| Label |
Cy5
|
| Label protocol |
200 ng of total RNA was labelled following the Agilent Two-color Low Input Quick Amp Labeling Protocol Version 6.5.
|
| |
|
| Channel 2 |
| Source name |
Bcl-xAS-transfected 786-O cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: 786-O
original tissue: clear cell renal cell cancer (CCRCC)
transfection: Bcl-xAS
|
| Treatment protocol |
Cells were transfected with 6 μg of Bcl-xAS expression construct or with 6 μg of empty vector (pCEP) as control using 3:1 FuGENE HD reagent (Promega). At 24 h after transfection, total RNA was extracted.
|
| Growth protocol |
786-O cells were cultivated in 147.8 cm2 cell culture plates with Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS. To perform four biological replicates, we cultivated four plates in parallel.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol reagent following the manufacturer's instructions. Total RNA was DNase-I-treated and purified using the Illustra RNAspin Mini kit (GE Healthcare) following the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
200 ng of total RNA was labelled following the Agilent Two-color Low Input Quick Amp Labeling Protocol Version 6.5.
|
| |
|
| |
| Hybridization protocol |
300 ng of cRNA was hybridized following the Agilent Two-color Low Input Quick Amp Labeling Protocol Version 6.5.
|
| Scan protocol |
Scanned on an Agilent G2565CA Microarray Scanner System.
|
| Description |
Bcl-xAS_rep2_Cy3/Ctl_rep2_Cy5
Bcl-xAS-transfected 786-O cells, biological replicate 2 of 4, Cy3 labelled vs. empty vector-transfected 786-O cells, biological replicate 2 of 4, Cy5 labelled.
|
| Data processing |
Linear and lowess-normalized data (Processed Signal Column by Feature Extractor Agilent Software v.10.7.1.1) was transformed to log2 ratios for test/reference samples.
|
| |
|
| Submission date |
Jun 17, 2012 |
| Last update date |
Mar 02, 2021 |
| Contact name |
Angela A. Fachel |
| E-mail(s) |
anf2032@med.cornell.edu
|
| Organization name |
Weill Cornell Medicine
|
| Street address |
1300 York Ave
|
| City |
New York |
| State/province |
New York |
| ZIP/Postal code |
10065 |
| Country |
USA |
| |
|
| Platform ID |
GPL13607 |
| Series (1) |
| GSE38766 |
Expression profile of human 786-O cells overexpressing Bcl-xAS lncRNA |
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