|
| Status |
Public on Jun 10, 2015 |
| Title |
GSM-005 |
| Sample type |
RNA |
| |
|
| Source name |
infected spleen
|
| Organism |
Microtus fortis |
| Characteristics |
tissue: spleen
Sex: Male
infection: schistosomes
|
| Treatment protocol |
All animals were singly-housed for one week before infection. Food and water was available ad libitum. M.fortis, Rat and BALB/c mice were percutaneously infected with 3000, 2000 and 200 S. japonicum cercariae (Chinese mainland strain, Anhui population), respectively. Three additional animals were used as uninfected controls.
|
| Growth protocol |
All animal care and experimental procedures were conducted according to the guidelines for animal use in toxicology.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The animals were euthanized at 10 days post-infection (p.i.) and the lungs, liver and spleen were harvested and preserved in RNA later (Ambion) at -80℃ until RNA extraction. Total RNA extraction from ~100mg tissues was performed with the mirVana isolation kit, according to instructions of the manufacturer (Ambion, USA). The quality of RNA was measured using a Nanodrop-1000 and the integrity was evaluated by Agilent 2100 Bioanalyzer (Agilent Technologies, USA ).
|
| Label |
cy5
|
| Label protocol |
The assay began with 5ug of RNA from each sample which was size fractionated using an YM-100 Microcon filter (Millipore, Bedford, MA, USA). The small RNAs (<300 nt) extracted were 3’-extended with poly(A) polymerase and labeled with Cy5 dye. Hybridizations were started according to the manual using a μParaflo@ microfluidic technology.
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| |
|
| Hybridization protocol |
The Cy5 dye labeled small RNAs (<300nt) were dissolved in 100 ul 6xSSPE buffer (0.90 M NaCl, 60 mM Na2HPO4, 6 mM EDTA, pH 6.8) containing 25% formamide at 34℃ overnight.
|
| Scan protocol |
Hybridization images were scanned using a laser scanner (GenePix 4000B, Molecular Device) and digitize analysis was performed using Array-Pro software (Media Cybernetics, Bethesda, MD).
|
| Description |
sample5 GSM-005
|
| Data processing |
The data was normalized using a cyclic LOWESS (Locally-weighted Regression) method for further analysis. We definded the differentially expressed miRNA using the ratio of detected signals log2-fold changes [log2(infected/control)] and p values of the t test were calculated.
|
| |
|
| Submission date |
Jun 19, 2012 |
| Last update date |
Jun 10, 2015 |
| Contact name |
hongxiao han |
| E-mail(s) |
hhx021@yahoo.com.cn
|
| Organization name |
Chinese Academy of Agricultural Sciences
|
| Department |
Shanghai Veterinary Research Institute
|
| Street address |
No. 518, Ziyue Road,
|
| City |
Shanghai |
| ZIP/Postal code |
200241 |
| Country |
China |
| |
|
| Platform ID |
GPL15710 |
| Series (1) |
| GSE38802 |
MicroRNA expression profiles associated with anti-schistosome features in Microtus fortis |
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