|
| Status |
Public on Jun 04, 2014 |
| Title |
shTET1#2 |
| Sample type |
RNA |
| |
|
| Source name |
HEK293T_TET1 shRNA
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HEK293T
transfected with: TET1 shRNA#2
|
| Treatment protocol |
mTET1-CD, TET1-CD, mTET1-FL, TET1-FL expression plasmid or empty vector was transfected into HEK293T cells by using OPTI-MEM (Invitrogen) and FuGene HD transfection reagent (Roche). Three days after transfection, overexpressed cells were purified with fluorescent activated cell sortng. Lentiviral shRNA-mediated TET1 knockdown were also established in HEK293T cells, and one control and two TET1 knockdown clones were identified.
|
| Growth protocol |
HEK293T cells were cultured in Dulbecco's Modified Eagle Medium modified with 10% fetal bovine serum and 100 μg/ml streptomycin-penicillin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
100 ng of total RNA were amplified using the Ambion WT Expression Kit and cDNA were biotin labelled using Affymetrix GeneChip WT Terminal Labeling protocol.
|
| |
|
| Hybridization protocol |
Affymetrix GeneChip Human Gene 2.0 ST arrays were hybridized to 5.5ug of biotin labeled cDNA, washed and stained according to Affymetrix protocol.
|
| Scan protocol |
GeneChips were scanned using GeneChip Scanner 3000 (Affymetrix)
|
| Description |
Sample 3
|
| Data processing |
The data were analyzed using the Affymetrix Gene Expression Console (v.1.3.1) with RMA (Robust Multi-array Average) normalization algorithm.
|
| |
|
| Submission date |
Aug 20, 2013 |
| Last update date |
Jun 04, 2014 |
| Contact name |
CHUNLEI JIN |
| E-mail(s) |
jincl1980@gmail.com
|
| Organization name |
The University of Texas MD Anderson Cancer Center
|
| Street address |
1515 Holcombe Blvd
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
| |
|
| Platform ID |
GPL16686 |
| Series (2) |
| GSE44039 |
TET1 is a maintenance DNA demethylase that prevents methylation spreading in adult cells |
| GSE50016 |
TET1 is a maintenance DNA demethylase that prevents methylation spreading in adult cells [cDNA microarray] |
|
