|
| Status |
Public on Dec 31, 2009 |
| Title |
ZNF191-overexpressed cells vs Control |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
human embryo kidney cells transfected with pcDNA3-ZNF191
|
| Organism |
Homo sapiens |
| Characteristics |
ZNF191-overexpressed cells
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using the NucleoSpin RNAII Kit (Macherey-Nagel, Düren, DE) and quantified via spectrophotometry (Nanodrop, Labtech). RNA integrity was confirmed using the Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA).
|
| Label |
cy3
|
| Label protocol |
Agilent’s recommended procedures for microarray-based one-color gene expression analysis were followed. Briefly, 350ng of total RNA with control RNA Spike In were amplified and labeled with Cy3-CTP using the T7-based Low RNA Input Linear Amplification Kit, PLUS, One-Color (Agilent Technologies, Santa Clara, CA). After purification (RNeasy Mini Kit, QIAGEN), labeled cRNAs were quantified for yield and dye incorporation using a Nanodrop spectrophotometer.
|
| |
|
| Channel 2 |
| Source name |
human embryo kidney cells transfected with pcDNA3
|
| Organism |
Homo sapiens |
| Characteristics |
HEK293 cell line
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using the NucleoSpin RNAII Kit (Macherey-Nagel, Düren, DE) and quantified via spectrophotometry (Nanodrop, Labtech). RNA integrity was confirmed using the Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA).
|
| Label |
cy5
|
| Label protocol |
Agilent’s recommended procedures for microarray-based one-color gene expression analysis were followed. Briefly, 350ng of total RNA with control RNA Spike In were amplified and labeled with Cy5-CTP using the T7-based Low RNA Input Linear Amplification Kit, PLUS, One-Color (Agilent Technologies, Santa Clara, CA). After purification (RNeasy Mini Kit, QIAGEN), labeled cRNAs were quantified for yield and dye incorporation using a Nanodrop spectrophotometer.
|
| |
|
| |
| Hybridization protocol |
1.65microg of Cy3-labeled RNA and Cy5-labeled RNA were hybridized to each array in a 110microl total volume of hybridization buffer (Agilent Technologies) for 17 hours at 65°C. Washing was performed according to the Agilent protocol.
|
| Scan protocol |
Arrays were analyzed using the dynamic autofocus Agilent G2565BA microarray scanner and the Agilent Feature Extraction software (FE v9.4.1, GE1-v5_91_0806 protocol).
|
| Description |
Poor quality spots were flagged according to default FE criteria and corresponding values were removed.
|
| Data processing |
GeneSpring 10.0 quantile normalization
|
| |
|
| Submission date |
Dec 30, 2008 |
| Last update date |
Jan 05, 2009 |
| Contact name |
Jianzhong Li |
| E-mail(s) |
lijianzhong1234@yahoo.com
|
| Organization name |
Second Military Medical University
|
| Department |
Department of Biochemical Pharmacy
|
| Street address |
GuoHe Road 325#
|
| City |
ShangHai |
| ZIP/Postal code |
200433 |
| Country |
China |
| |
|
| Platform ID |
GPL6480 |
| Series (1) |
| GSE14273 |
The target genes of transcript factor ZNF191 |
|
