|
| Status |
Public on May 10, 2019 |
| Title |
Uninfected hepatocytes 8.1 |
| Sample type |
RNA |
| |
|
| Source name |
Primary hepatocyte culture, uninfected
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Hepatocytes
infection: uninfected
|
| Treatment protocol |
Co-incubated with freshly isolated Py-GFP sporozoites from mosquitoes
|
| Growth protocol |
Primary hepatocyte cultures maintained in 6 well-plates
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The hepatocytes were detached by trypsinization (0.25% Trypsin-EDTA), FACS sorted directly into fetal calf serum. The hepatocytes were pelleted and immediately used for RNA extraction. Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
Biotin
|
| Label protocol |
100 ng of total RNA was converted to amplified ss-cDNA using the GeneChip WT Plus Reagent Kit (Affymetrix,Cat. #902513). Following fragmentation (fragment size ~40-70nt ), the samples were labeled with biotin.
|
| |
|
| Hybridization protocol |
Biotin-labeled ss-cDNA was then mixed with Affymetrix eukaryotic hybridization buffer, placed onto the Affymetrix GeneChip Mouse Transcriptome Array 1.0 (Part No. 902511), and incubated at 45ยบ C for 18 h with 60 rpm rotation
|
| Scan protocol |
GeneChips were scanned using the Affymetrix Model 3000 scanner (with 7G upgrade).
|
| Description |
Gene expression in uninfected hepatocytes
|
| Data processing |
The data were collected using Affymetrix Comand Console operating software v4.1.2 and Affymetrix Expression Console Software Build 1.4.4.46 using Affymetrix default analysis settings and global scaling as normalization method.
|
| |
|
| Submission date |
May 09, 2019 |
| Last update date |
May 10, 2019 |
| Contact name |
Samarchith Kurup |
| E-mail(s) |
[email protected]
|
| Organization name |
The University of Iowa
|
| Street address |
51 Newton Rd
|
| City |
Iowa city |
| State/province |
Iowa |
| ZIP/Postal code |
52242 |
| Country |
USA |
| |
|
| Platform ID |
GPL20258 |
| Series (1) |
| GSE130961 |
Comparing transcrional changes in Plasmodium yoelii infected to uninfected primary hepatocytes |
|
