|
| Status |
Public on Dec 31, 2012 |
| Title |
KB PST 2 |
| Sample type |
RNA |
| |
|
| Source name |
KB +PST
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: KB
cell type: Nasopharyngeal tumor
|
| Treatment protocol |
Cells were treated with PST001 polysaccharide along with a control.
|
| Growth protocol |
A549, KB and MCF-7 cells were collected when they attains 70% of cell growth. Removed the growth medium by aspiration and washed cells once with 2 ml of phosphate buffered saline. Washed the cells again with 1 ml of PBS-EDTA. Trypsinized cells for 3-5 minutes in 0.5 ml of 0.25% trypsin at 37°C. Diluted the cells with 1 ml of growth medium to inactivate the trypsin. Transferred 1.5 ml of the cell suspension to a 15 ml falcon tube and centrifuged at 2500 rpm for 10 min. Supernatent was discarded and pellet was transferred to fresh T25 tissue culture flask and added 5 ml fresh growth medium. Placed the cells in a 37°C incubator at 5% CO2 and monitored cell-density daily.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by Trizol method. RNA was cleaned up using RNeasy MinElute Cleanup Kit (Qiagen) and reverse transcribed.
|
| Label |
Biotin
|
| Label protocol |
The product cDNA is converted into cRNA by T7 polymerase and labelled with Cy3 (labelled on cytosine) using Agilent kit.
|
| |
|
| Hybridization protocol |
RNA was fragmented by metallic cleavage using manufacturar's protocol. Array lanes were blocked using ssDNA and subsequently samples were hybridized at 65oC for 17h. Array was washed with wash buffer with and without DNA. Finally array was dried.
|
| Scan protocol |
Arrays were scanned on Illumina iScan system.
|
| Data processing |
Initial data was extracted by gene-expression module of GenomeStudio version 2010.2. Data was analyzed by Genespring GX12.0 software.
|
| |
|
| Submission date |
Oct 15, 2012 |
| Last update date |
Dec 31, 2012 |
| Contact name |
madavan vasudevan |
| E-mail(s) |
madavan.vasudevan@gmail.com
|
| Phone |
+91-9845093355
|
| Organization name |
Theomics International Private Limited
|
| Street address |
Kasturi Nagar
|
| City |
Bangalore |
| State/province |
Karnataka |
| ZIP/Postal code |
560043 |
| Country |
India |
| |
|
| Platform ID |
GPL10904 |
| Series (1) |
| GSE41598 |
Gene Expression analysis of PST001 in cancer cell lines |
|
