|
| Status |
Public on Aug 17, 2019 |
| Title |
Peripheral Blood Mononuclear Cells, Healthy control 33 |
| Sample type |
RNA |
| |
|
| Source name |
Peripheral Blood Mononuclear Cells
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: whole blood
gender: male
|
| Treatment protocol |
Overnight fasting blood samples were drawn by venipuncture from all patients and stored at -80 ℃ prior to use. PBMCs were isolated from the middle white monolayer by density gradient centrifugation using lymphocyte separation medium (TBD, Tianjin, China).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the TRIzol reagent following the manufacturer's recommendations. RNA was quantified using a NanoDrop-2000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
14μl cDNA labeled with a 1μl fluorescent dye (Cy3-dCTP) was produced by Eberwine’s linear RNA amplification method and subsequent enzymatic reaction. This procedure has been previously described, and the procedure has been improved by using the WT Expression Kit (No. A24696C, Thermo Fisher Scientific)for producing higher yields of labeled cDNA.
|
| |
|
| Hybridization protocol |
cDNA in hybridization solution was denatured at 95℃ for 3 min prior to loading onto a microarray. Human lncRNA+mRNA Array V4.0 (4×180 K) were hybridized was preformed in a Agilent Hybridization Oven overnight at a rotation speed of 20 rpm at 45℃. After hybridization, microarrays were washed 6 minutes at 42℃ with GE Wash Buffer 1 (0.2% SDS，2×SSC) and 4 minutes with 42°C GE Wash buffer 2 (0.2% SDS，2×SSC), then dried immediately by brief centrifugation.
|
| Scan protocol |
Scanned on an Agilent G2565CA scanner.
|
| Description |
Gene expression
|
| Data processing |
Feature Extraction v10.7 (Agilent Technologies, CA) software was used to extract all features of the data obtained from the scanned images and Agilent genespring software was used to analyze the raw data, which are normalized by percentile normalization.
|
| |
|
| Submission date |
Apr 12, 2018 |
| Last update date |
Aug 17, 2019 |
| Contact name |
Lin Li |
| E-mail(s) |
lilinshine@163.com
|
| Organization name |
Department of Epidemiology, State Key Laboratory of Cardiovascular Disease
|
| Department |
Department of Epidemiology
|
| Lab |
State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Diseases
|
| Street address |
Xicheng District, No.167 Beilishi Road
|
| City |
Beijing |
| State/province |
Beijing |
| ZIP/Postal code |
100037 |
| Country |
China |
| |
|
| Platform ID |
GPL20115 |
| Series (1) |
| GSE113079 |
LncRNA Expression Profile and Identification of Novel LncRNA Biomarkers for Diagnosing Coronary Artery Disease |
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