|
| Status |
Public on May 31, 2011 |
| Title |
Huh7 EZH2 ChIP-chip Replicate 1 Slide 1 |
| Sample type |
genomic |
| |
|
| Channel 1 |
| Source name |
Huh7 cells immunoprecipitated
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Huh7
|
| Treatment protocol |
No treatment was performed. EZH2 target genes in normal condition were identified.
|
| Growth protocol |
The 2 HCC cell lines were maintained in DMEM supplemented with 10% FBS.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
ChIP assays were performed as described previously (Lee et al. Nat Protoc 2006;1:729-748). IP and input DNA were then purified with standard phenol/chloroform method.
|
| Label |
Cy5
|
| Label protocol |
DNA were amino-allyl labelled using randomly-primed, Klenow-based extension protocol. Cy5/3 dye was then incorporated into the DNA using in-direct labeling by incubating the dye with DNA for 3 hours.
|
| |
|
| Channel 2 |
| Source name |
Huh7 cells input
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: Huh7
|
| Treatment protocol |
No treatment was performed. EZH2 target genes in normal condition were identified.
|
| Growth protocol |
The 2 HCC cell lines were maintained in DMEM supplemented with 10% FBS.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
ChIP assays were performed as described previously (Lee et al. Nat Protoc 2006;1:729-748). IP and input DNA were then purified with standard phenol/chloroform method.
|
| Label |
Cy3
|
| Label protocol |
DNA were amino-allyl labelled using randomly-primed, Klenow-based extension protocol. Cy5/3 dye was then incorporated into the DNA using in-direct labeling by incubating the dye with DNA for 3 hours.
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (Agilent In Situ Hybridization Kit Plus) were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequential
|
| Scan protocol |
Scanned on an Agilent G2505B scanner.
Images were quantified using Agilent Feature Extraction Software (version 10.5.1.1).
|
| Description |
Huh7 Replicate 1 of 2
|
| Data processing |
Agilent Feature Extraction Software (10.5.1.1) was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Aug 20, 2009 |
| Last update date |
May 31, 2011 |
| Contact name |
Richard KW CHOY |
| E-mail(s) |
[email protected]
|
| Phone |
852-26323099
|
| Fax |
852-26360008
|
| Organization name |
The Chinese University of Hong Kong
|
| Department |
Obstetrics & Gynaecology
|
| Street address |
1/F, Block E, Prince of Wales Hospital
|
| City |
Shatin |
| ZIP/Postal code |
SAR |
| Country |
Hong Kong |
| |
|
| Platform ID |
GPL4124 |
| Series (1) |
| GSE17733 |
Delineation of EZH2 oncogenic functions in hepatocellular carcinoma |
|
