|
| Status |
Public on Dec 05, 2019 |
| Title |
H1299-con sh |
| Sample type |
RNA |
| |
|
| Source name |
non small lung cancer cell
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: H1299
cell type: non small lung cancer cell
tissue: lung
genotype/variation: control
|
| Treatment protocol |
CNOT2 gene was knocked down for 48 hr and melatonin 1mM was treated for 24 hr.
|
| Growth protocol |
A549 and H1299 were grown in RPMI with 10% FBS and antibiotics. XO1 and XO2 were grown in DMEM-F12 with B27 and antibiotics supplemented with growth factors (EGF and bFGF).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
Amplified and labeled cRNA was purified on cRNA Cleanup Module (Agilent Technology) according to the manufacturer’s protocol. Labeled cRNA target was quantified using ND-1000 spectrophotometer (NanoDrop Technologies, Inc., Wilmington, DE).
|
| |
|
| Hybridization protocol |
After checking labeling efficiency, fragmentation of cRNA was performed by adding 10X blocking agent and 25X fragmentation buffer and incubating at 60oC for 30 min. The fragmented cRNA was resuspended with 2X hybridization buffer and directly pipetted onto assembled Agilent’s Human Oligo Microarray (60K). The arrays hybridized at 65oC for 17 hours using Agilent Hybridization oven (Agilent Technology, USA). The hybridized microarrays were washed as the manufacturer’s washing protocol (Agilent Technology, USA).
|
| Scan protocol |
The hybridized images were scanned using Agilent’s DNA microarray scanner and quantified with Feature Extraction Software (Agilent Technology, Palo Alto, CA).
|
| Description |
3. H1299-con sh
|
| Data processing |
All data normalization and further analysis were performed using GeneSpringGX 7.3 (Agilent Technology, USA).
|
| |
|
| Submission date |
Dec 20, 2016 |
| Last update date |
Dec 05, 2019 |
| Contact name |
Hyemin Lee |
| E-mail(s) |
glansy555@gmail.com
|
| Organization name |
Kyunghee Univ
|
| Street address |
Kyunghee-dae-ro 26
|
| City |
Seoul |
| ZIP/Postal code |
131701 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE92612 |
CNOTshRNA or Melatonin treated samples |
|
