|
| Status |
Public on Aug 01, 2016 |
| Title |
296-4 |
| Sample type |
RNA |
| |
|
| Source name |
Jurkat cells co-cultured with ionomycin and phorbol 12-myristate 13-acetate
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Jurkat cells
treatment: co-cultured with ionomycin and phorbol 12-myristate 13-acetate
|
| Treatment protocol |
Jurkat cells co-cultured with 10 ng/mL phorbol 12-myristate 13-acetate (PMA) and 500 ng/mL ionomycin 24 h at 37ºC.
|
| Growth protocol |
Jurkat cells (1×106/mL) were cultured in RPMI-1640 containing heat-inactivated fetal bovine serum (10%), L-glutamine (2 mmol/L), penicillin (100 U/mL) and streptomycin (100 mg/mL)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from purified T cells or Jurkat cells using the Quick-RNA MiniPrep kit (Zymo Research, Irvine, CA, USA) according to the manufacturer’s protocol. The RNA concentration was quantified using a spectrophotometer (NanoDrop 1000, Thermo Fisher Scientific, Waltham, MA, USA).
|
| Label |
Cy3
|
| Label protocol |
0.2 μg of total RNA was amplified by a Low Input Quick-Amp Labeling kit (Agilent Technologies, USA) and labeled with Cy3 (CyDye, Agilent Technologies, USA) during the in vitro transcription process.
|
| |
|
| Hybridization protocol |
0.6 μg of Cy3-labled cRNA was fragmented to an average size of about 50-100 nucleotides by incubation with fragmentation buffer at 60°C for 30 minutes. Correspondingly fragmented labeled cRNA is then pooled and hybridized to Agilent SurePrint G3 Human V2 GE 8×60K Microarray (Agilent Technologies, USA) at 65°C for 17 h.
|
| Scan protocol |
Scanned images are analyzed by Feature extraction10.5.1.1 software (Agilent Technologies, USA),
|
| Description |
Gene expression after 24hr in Jurket cells co-cultured with ionomycin and phorbol 12-myristate 13-acetate
|
| Data processing |
An image analysis and normalization software is used to quantify signal and background intensity for each feature.
|
| |
|
| Submission date |
Aug 14, 2015 |
| Last update date |
Aug 01, 2016 |
| Contact name |
Hui Chun Yu |
| E-mail(s) |
junvsusagi@gmail.com
|
| Phone |
+886920179628
|
| Organization name |
Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation
|
| Department |
Medical research
|
| Street address |
No. 2, Min-Sheng Rd, Dalin Town
|
| City |
Chia-Yi |
| ZIP/Postal code |
62247 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE72100 |
Increased expression of long non-coding RNAs in T cells from patients with rheumatoid arthritis facilitates the inflammatory responses |
|
