|
| Status |
Public on Mar 08, 2018 |
| Title |
T_HC_C2 [Human LncRNA Array v3.0] |
| Sample type |
RNA |
| |
|
| Source name |
T cell, HC
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Peripheral Blood Mononuclear Cells
gender: female
|
| Treatment protocol |
CD3+ T cell was positively labeled with anti-human CD3 magnetic Particles-DM (BD IMag™, San Jose, USA) and selected by IMag magnetic cell separation system (BD IMagnet™, San Jose, USA).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from T cells by using TRIzol Reagent (Invitrogen, Carlsbad, USA) according to the manufacture’s protocol. The purity and concentration of RNA was evaluated with the NanoDrop ND-1000 (Thermo Fisher Scientific, Wilmington, DE).
|
| Label |
Cy3
|
| Label protocol |
Each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 3’ bias utilizing a random priming method. The concentration and specific activity of the labeled cRNAs (pmol Cy3/μg cRNA) were measured by NanoDrop ND-1000.
|
| |
|
| Hybridization protocol |
1 μg of each labeled cRNA was fragmented by adding 5 μl 10 × Blocking Agent and 1 μl of 25 × Fragmentation Buffer, then heated the mixture at 60 °C for 30 min, finally 25 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 50 μl of hybridization solution was dispensed into the gasket slide and assembled to the LncRNA expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven
|
| Scan protocol |
The Agilent Scanner G2505B was used to scan the slides
|
| Description |
lncRNA expression in T cells from healthy volunteer C2
|
| Data processing |
Agilent Feature Extraction software was applied to asses the array images. Quantile normalization and subsequent data processing were performed with the GeneSpring GX v11.5.1 software package (Agilent Technologies).After quantile normalization of the raw data, LncRNAs and mRNAs that at least 3 out of 9 samples have flags in Present or Marginal (“All Targets Value”) were chosen for further data analysis.Differentially expressed LncRNAs and mRNAs with statistical significance were identified through Volcano Plot filtering.
|
| |
|
| Submission date |
Jul 21, 2016 |
| Last update date |
Mar 08, 2018 |
| Contact name |
Lian-Ju Li |
| Organization name |
Anhui Medical University
|
| Department |
Department of Epidemiology and Biostatistics, School of Public Health
|
| Street address |
81 Meishan Road
|
| City |
Hefei |
| State/province |
Anhui |
| ZIP/Postal code |
230032 |
| Country |
China |
| |
|
| Platform ID |
GPL16956 |
| Series (2) |
| GSE84654 |
Comprehensive lncRNAs expression profiling reveals the potential role of lncRNAs in systemic lupus erythematosus [Human LncRNA Array v3.0] |
| GSE84656 |
Comprehensive expression profiling reveals the potential role of lncRNAs and circRNAs in systemic lupus erythematosus |
|
