|
| Status |
Public on May 07, 2009 |
| Title |
LATENT_PBMC_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
latent infected, PBMCs, pool of 4 individuals
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Peripheral Blood Mononuclear Cells
gender: two males and two females
age: 30y～45y
|
| Treatment protocol |
The peripheral blood mononuclear cells were freshly extracted with the human lymphocyte separation medium from the peripheral blood drawn from the volunteers of TB patients, latent-infected individuals and the completely healthy persons, respectively.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was prepared using the TRizol LS reagent (Invitrogen, CA, USA ) following the manufacturer's recommendations. RNA was quantified using spectrophotometer method and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
The microRNAs were labled using the miRNA Complete Labeling and Hyb Kit. Details were that 1005ng total RNA was treated with phosphatase at 37 ℃for 30min. The dephosphorylated RNA was labled as the kit instruction .
|
| |
|
| Hybridization protocol |
The Cy3-labled RNA was dried for 1-2h at 45℃ to 55℃, then assembled the hybridization mixture and hybridize for 20h at 55℃. Finally the hibridized slides were washed for scanning
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent Microarray Scanner (G2565BA) using one color scan setting for 1x44k array slides [Scan Area 61x21.6 mm, Scan resolution 5um, Dye channel is set to Green and Green PMT is set to 100% (XDR Hi) and 5%(XDR Lo)].
|
| Description |
microRNA level in PBMCS of the latent-infected individuals
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.5 (Agilent G4450AA) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
May 06, 2009 |
| Last update date |
Jan 29, 2010 |
| Contact name |
Zhaogang Sun |
| E-mail(s) |
sunzg75@hotmail.com
|
| Phone |
86-10-69545354
|
| Fax |
86-10-69546819
|
| Organization name |
Beijing Tuberculosis & Thotacic Tumor research Institute
|
| Department |
Microbiology and Immunology
|
| Lab |
Prof. Li's lab
|
| Street address |
No. 97, Machang Road, Tongzhou District
|
| City |
Beijing |
| State/province |
Beijing |
| ZIP/Postal code |
101149 |
| Country |
China |
| |
|
| Platform ID |
GPL8227 |
| Series (1) |
| GSE15977 |
Analysis of the microRNA that involved the tuberculosis or latent TB infection I |
|
