|
| Status |
Public on Nov 23, 2011 |
| Title |
Control 3 [mRNA] |
| Sample type |
RNA |
| |
|
| Source name |
PBMC
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Blood
infection: none
|
| Treatment protocol |
One half of the cells were subsequently infected with 0.1 MOI of virus particles using standard protocols as described and the remaining half of the culture is maintained under similar conditions and used as control. Seven days post infection (pi), cells were collected and frozen for total RNA isolation.
|
| Growth protocol |
We purchased normal donor blood from the American Red Cross Blood Bank in Pittsburgh using appropriate IRB approval forms from the University of Pittsburgh. PBMCs were isolated by Ficoll-Hypaque gradient centrifugation. Freshly isolated normal donor PBMCs (5×106/mL) were stimulated with 5 µg/ml PHA-P (Sigma, St. Louis, MO) for three days. Cells were washed, divided into two parts and cultured in RPMI medium (GIBCO, CA) containing 10% FBS (Hyclone, Logan, UT), 1% L-glutamine (Cambrex, MD), 1% penicillin-streptomycin (GIBCO, CA), and IL-2 (200 U/mL, Chiron, Emoryville, CA).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using the TRIzol method (InVitrogen) as suggested by the manufacturer.
|
| Label |
biotin
|
| Label protocol |
Total RNA samples were labeled using ‘TotalPrep RNA’ labeling kit (Ambion)
|
| |
|
| Hybridization protocol |
hybridized onto array bead chips overnight on rocker
|
| Scan protocol |
scanned on ‘iScan system’ according to the manufacturer's protocols
|
| Description |
Uninfected PBMC from donor 3
|
| Data processing |
Initial raw data analysis and cubic spline normalization was done using BeadStudio Gene expression module (Illumina, Inc.).
|
| |
|
| Submission date |
Nov 22, 2011 |
| Last update date |
Nov 23, 2011 |
| Contact name |
Pruthvi S. R. Nagilla |
| E-mail(s) |
prn9@pitt.edu
|
| Phone |
412 624-3062
|
| URL |
http://www.idm.pitt.edu/directory/bios/ayyavoo.asp
|
| Organization name |
University of Pittsburgh
|
| Department |
Infectious Diseases and Microbiology
|
| Street address |
416 Parran Hall, 130 DeSoto Street
|
| City |
Pittsburgh |
| State/province |
PA |
| ZIP/Postal code |
15261 |
| Country |
USA |
| |
|
| Platform ID |
GPL6947 |
| Series (2) |
| GSE33877 |
Comparative Expression Profile of miRNA and mRNA in Primary Peripheral Blood Mononuclear Cells Infected with Human Immunodeficiency Virus (HIV-1) [mRNA] |
| GSE33879 |
Comparative Expression Profile of miRNA and mRNA in Primary Peripheral Blood Mononuclear Cells Infected with Human Immunodeficiency Virus (HIV-1) |
|
