|
| Status |
Public on Mar 11, 2010 |
| Title |
ILTV Infected chicken embryo lung cells at 3 dpi Replicate 3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Embryo lung cells
|
| Organism |
Gallus gallus |
| Characteristics |
infection: none
cell type: embryo lung cells
|
| Growth protocol |
Cells were grown in a 1:1 ratio of mammary epithelial growth media (MEGM; Lonza, Rockland, ME, USA) and Dulbecco’s Modified Eagle’s Medium (DMEM, 0.45% glucose) plus 2% fetal bovine serum (FBS), 100 units/ml penicillin, 100 µg/ml streptomycin, and 2 mM L-glutamine in 10 cm tissue culture plates (Sarstedt Inc., Newton, NC, USA) pretreated with 0.5 % gelatin in PBS to improve cell adhesion.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
2 µg of total RNA were used for this experiment, and all procedures were followed to manufacturer's instructions.
|
| |
|
| Channel 2 |
| Source name |
Embryo lung cells
|
| Organism |
Gallus gallus |
| Characteristics |
infection: infectious laryngotracheitis virus (ILTV)
cell type: embryo lung cells
time point: 3 dpi
replicate: 3
|
| Treatment protocol |
ILTV was infected the chicken embryonic lung cells at a multiplicity of infection (m.o.i.) of 0.1.
|
| Growth protocol |
Cells were grown in a 1:1 ratio of mammary epithelial growth media (MEGM; Lonza, Rockland, ME, USA) and Dulbecco’s Modified Eagle’s Medium (DMEM, 0.45% glucose) plus 2% fetal bovine serum (FBS), 100 units/ml penicillin, 100 µg/ml streptomycin, and 2 mM L-glutamine in 10 cm tissue culture plates (Sarstedt Inc., Newton, NC, USA) pretreated with 0.5 % gelatin in PBS to improve cell adhesion.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
2 µg of total RNA were used for this experiment, and all procedures were followed to manufacturer's instructions.
|
| |
|
| |
| Hybridization protocol |
An equal amount (825 ng) of Cy3 and Cy5 labeled cRNA probes were mixed with gene expression hybridization kit (Agilent, part No. 5188-5242) and hybridized on a 4X44K Agilent custom chicken oligo microarray (array ID: 017698). The hybridized slides were washed using a commercial kit package (Agilent).
|
| Scan protocol |
Scanned on an Genepix 4000B scanner (Molecular Devices Corporation)
|
| Description |
Biological replicate 3 of 3. ILTV infected chicken embryo lung cells, harvested at 3dpi.
|
| Data processing |
Gene Pix Pro (v 6.1) with Accuity software (v 4.0). Background-corrected red and green intensities for each spot were used in the subsequent analysis. Global normalization based on local polynomial regression (loess) was applied to the intensities for removing effects which arise from undesirable systematic variations in microarray experiment.
|
| |
|
| Submission date |
Mar 04, 2010 |
| Last update date |
Mar 10, 2010 |
| Contact name |
Byung-whi Kong |
| E-mail(s) |
bkong@uark.edu
|
| Phone |
479-575-5494
|
| Organization name |
University of Arkansas
|
| Department |
Poultry Science
|
| Street address |
1260 W. Maple, POSC, L-426
|
| City |
Fayetteville |
| State/province |
AR |
| ZIP/Postal code |
72701 |
| Country |
USA |
| |
|
| Platform ID |
GPL6413 |
| Series (1) |
| GSE20630 |
Transcriptional Profiling of Host Gene Expression in Chicken Embryo Lung Cells Infected with Laryngotracheitis Virus |
|
