|
| Status |
Public on Mar 14, 2009 |
| Title |
PBMCs_with_malaria_symptoms_patient_06 |
| Sample type |
RNA |
| |
|
| Source Name |
Peripheral Blood Mononuclear Cells from patients with malaria symptons
|
| Organism(s) |
Homo sapiens |
| Characteristics |
gender: Male
age: 30
symptoms: PBMC in parasitemic, symptomatic (fever, headache, etc) individual prior to treatment
|
| Treatment protocol |
Cells were immediately frozen at -80°C in RNeasy extraction buffer (Qiagen).
|
| Growth protocol |
Peripheral Blood Mononuclear Cells (PBMCs) were isolated from undiluted whole blood on Ficoll-paque Plus (GE Healthcare) per the manufacturer’s recommendations. After isolation, PBMCs were washed in PBS and frozen as described below.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from PBMCs using an RNeasy kit (Qiagen) following the manufacturer’s recommendations. RNA integrity and purity was analyzed with an Experion Automated Electrophoresis System (Bio-Rad) and a ND-1000 spectrophotometer (NanoDrop Technologies Inc.).
|
| Label |
Cy3
|
| Label protocol |
300 ng of total RNA was reverse transcribed and in vitro transcribed to cRNA using the Illumina TotalPrep RNA Amplification Kit (Illumina) according to the manufacturer's protocol
|
| |
|
| Hybridization protocol |
The cRNA (1.5 µg) was mixed with hybridization buffer and added to the BeadChips for hybridization at 58°C for 18 hours according to the manufacturer's protocol. BeadChips were then washed, blocked, and stained with streptavidin-Cy3, before they were washed again and dried by centrifugation.
|
| Scan protocol |
The BeadChips were scanned on the Illumina BeadStation according to the manufacturer's protocol
|
| Description |
none
|
| Data processing |
Raw signal intensities were log2 transformed and cubic spline normalized in BeadStudio software v 3.2
|
| |
|
| Submission date |
Mar 13, 2009 |
| Contact name |
Harry Björkbacka |
| E-mail(s) |
harry.bjorkbacka@med.lu.se
|
| URL |
http://www.med.lu.se/expcard
|
| Organization name |
Lund University
|
| Department |
Dept. of Clinical Sciences, Malmö
|
| Lab |
Experimental Cardiovascular Research Unit
|
| Street address |
CRC 91:12, UMAS entrance 72
|
| City |
Malmö |
| ZIP/Postal code |
205 02 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL6102 |
| Series (1) |
| GSE15221 |
Malaria primes the innate immune response due to IFNg induced enhancement of Toll-like receptor expression and function. |
|
