|
| Status |
Public on Nov 16, 2012 |
| Title |
3# sheep 3 dpi |
| Sample type |
RNA |
| |
|
| Source name |
3# H. contortus infected-sheep T lymphocytes, 3 dpi
|
| Organism |
Ovis aries |
| Characteristics |
gender: female
individual: 3# sheep
infectious agent: Haemonchus contortus
tissue: T lymphocytes
|
| Treatment protocol |
Peripheral blood samples from Haemonchus contortus infected sheep were collected in sodium citrate coated tubes at 0, 3-5, 25-30 and 60 days post-challenge (34) and were equally mixed with Hanks. Miscible liquids were slowly added above the isometric lymphocyte separation medium (Huadong Medicine). T Lymphocytes were then separated by centrifugation at 2000 g for 25 min, washed twice in Hanks and re-suspended in TRIzol reagent (Invitrogen).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The total RNA from the lymphocyte samples was obtained using TRIzol reagent (Invitrogen) following the manufacturer’s instructions and further purified using an RNeasy Mini kit (Qiagen). The quantity and quality of acquired RNA was measured using an Infinigen SSP-3300 ultramicrospectrophotometer and an Agilent 2100 Bioanalyzer.
|
| Label |
Cy3
|
| Label protocol |
Cy3 (GE healthcare) was labeled with synthesized cRNA using Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, , and purified using Rneasy Mini kit (Qiagen).
|
| |
|
| Hybridization protocol |
Labeled cRNA were hybridized to Agilent Whole Sheep Genome Oligo Microarrays according to the protocols provided by the manufacturer. After hybridization, microarrays were washed with GE Wash Buffer 1 (Agilent) and GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Microarrays were scanned by Agilent scanistor, scan resolution 5μm, PMT 100%.
|
| Description |
Gene expression at 3 days post infection in 3# Haemonchus contortus infected-sheep T lymphocytes
|
| Data processing |
Feature Extraction software was used to read the raw data. The raw data files were processed using the Agi4x44PreProcess package based on linear models for microarray data (limma) package developed within the Bioconductor project in the R statistical programming environment. Background correction and normalization, probes filtering and replicated probes merger were included in data preprocessing to generate microarray results for gene expression profiling.
|
| |
|
| Submission date |
Nov 15, 2012 |
| Last update date |
Nov 16, 2012 |
| Contact name |
Aifang Du |
| E-mail(s) |
afdu@zju.edu.cn
|
| Phone |
+86 571 8898 2583
|
| Fax |
+86 571 8898 2583
|
| Organization name |
Institute of Preventive Veterinary Medicine
|
| Department |
College of Animal Sciences, Zhejiang University
|
| Street address |
Yuhangtang Road 866
|
| City |
Hangzhou |
| State/province |
Zhejiang |
| ZIP/Postal code |
310058 |
| Country |
China |
| |
|
| Platform ID |
GPL16283 |
| Series (1) |
| GSE42302 |
Profiling of differentially expressed genes in sheep T lymphocytes response to Haemonchus contortus infection |
|
