|
| Status |
Public on Jan 05, 2010 |
| Title |
day 2-3 vs. sham |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
rat, AMI , 2days
|
| Organism |
Mus musculus |
| Characteristics |
tissue: myocardium in border zone
state: ischemia myocardium
strain: Sprague Dawley
|
| Treatment protocol |
The myocardial infarction (MI) models were induced by ligation of anterior descending coronary artery in adult Sprague-Dawley rats(180-200gram) as described by Shyu et al. On the 1st postoperative day , the survivals were randomly divided into three groups; 2days after MI group, 7 days after AMI group, 14days after AMI groups . These rat were exposed to all surgical procedures except the ligation of LAD artery formed into sham-control group.each group has five individal animals
|
| Growth protocol |
male sprague-Dawley rats were kept and bred in facilities of a standard conforming to UK home office requirements.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Rats were sacrificed with an overdose of pentobarbital 2, 7 or 14 days after myocardial infarction . Myocardium in border- zone were immediately snap frozen in liquid nitrogen for microarray analysis. Total RNA from rat myocardium in the border zone was isolated using trizol reagent (invitrogen, USA) according to manufacturer's directions.
|
| Label |
Cy3,Cy5
|
| Label protocol |
4μg of low-molecular-weight RNA was labeled with 500 ng 5'-phosphate-cytidyl-uridyl-cy3-3' (Dharmacon) with 2 units T4 RNA ligase (NEB). The labeling reaction was performed at 4℃ for 2 hours.
|
| |
|
| Channel 2 |
| Source name |
sham-control rat
|
| Organism |
Mus musculus |
| Characteristics |
tissue: myocardium
state: mixed normal myocardium from all sham-control rat
strain: Sprague Dawley
|
| Treatment protocol |
The myocardial infarction (MI) models were induced by ligation of anterior descending coronary artery in adult Sprague-Dawley rats(180-200gram) as described by Shyu et al. On the 1st postoperative day , the survivals were randomly divided into three groups; 2days after MI group, 7 days after AMI group, 14days after AMI groups . These rat were exposed to all surgical procedures except the ligation of LAD artery formed into sham-control group.each group has five individal animals
|
| Growth protocol |
male sprague-Dawley rats were kept and bred in facilities of a standard conforming to UK home office requirements.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Rats were sacrificed with an overdose of pentobarbital 2, 7 or 14 days after myocardial infarction . Myocardium in border- zone were immediately snap frozen in liquid nitrogen for microarray analysis. Total RNA from rat myocardium in the border zone was isolated using trizol reagent (invitrogen, USA) according to manufacturer's directions.
|
| Label |
Cy5,Cy3
|
| Label protocol |
4μg of low-molecular-weight RNA was labeled with 500 ng 5'-phosphate-cytidyl-uridyl-cy3-3' (Dharmacon) with 2 units T4 RNA ligase (NEB). The labeling reaction was performed at 4℃ for 2 hours.
|
| |
|
| |
| Hybridization protocol |
standard CapitalBio Methods: Labeled RNA was precipitated with 0.3 M sodium acetate, 2.5 volumes ethanol and resuspended in 15ul of hybridization buffer containing 3×SSC, 0.2% SDS and 15% formamide. The array was hybridized at 42℃ overnight and washed with two consecutive washing solutions (0.2% SDS, 2×SSC at 42°C for 5 min, and 0.2% SSC for 5 min at room temperature).
|
| Scan protocol |
Luxscan 10K/A (CapitalBio)
|
| Description |
compare miRNA expression profiles in border-zone myocardium in rat with acute myocardial infarction with myocardium in sham-control rat
|
| Data processing |
Intensity data was extracted from the TIFF images using LuxScanTM 3.0 software (CapitalBio Corp, Peking, China). The low-intensity spots were removed for which fewer than 30% of the signal pixels exceeded the median background plus two times its standard deviation.
|
| |
|
| Submission date |
Dec 29, 2009 |
| Last update date |
Jan 04, 2010 |
| Contact name |
bing shi |
| E-mail(s) |
dr_shibing@bjmu.edu.cn
|
| Organization name |
peking university third hospital
|
| Department |
department of cardiology
|
| Street address |
NO 39 huayuan north road
|
| City |
peking |
| ZIP/Postal code |
100191 |
| Country |
China |
| |
|
| Platform ID |
GPL9837 |
| Series (1) |
| GSE19695 |
Altered expression of microRNAs in the myocardium of rats with acute myocardial infarction |
|
