|
| Status |
Public on Aug 01, 2016 |
| Title |
PBMC_control_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
PBMC, healthy infant, replicate 2
|
| Organism |
Homo sapiens |
| Characteristics |
diagnosis: healthy
tissue: whole blood
cell type: PBMC
|
| Growth protocol |
PBMC was isolated by density gradient centrifugation on Percoll (d = 1.077) .
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions and purified by RNeasy MinElute Cleanup kit.
|
| Label |
Cy3
|
| Label protocol |
Complementary DNA amplification was carried out from 35 ng of total RNA using an Ovation PicoSL WTA System V2. After amplification, cDNA samples were labeled with Cy3 using Klenow polymerase.
|
| |
|
| Hybridization protocol |
Slide was hybridized with 1.65 μg Cy3-labeled cDNA using a Gene Expression Hybridization Kit (Agilent technologies, Santa Clara, CA, USA) in a hybridization oven.
|
| Scan protocol |
After 17 h of hybridization at 65 °C, the slide was washed with a Gene Expression Wash Buffer Kit (Agilent technologies, Santa Clara, CA, USA), according to the manufacturer’s instructions. Slides were scanned after washing on the Agilent DNA Microarray Scanner (G2505C) using one color scan setting for 8x60k array slides.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 11.0.1.1 (Agilent) using default parameters to obtain background subtracted and spatially detrended Processed Signal intensities. The intensity signals was imported to CLC Main workbench 6.0 and normalized by the quartile normalization method and log2 transformed.
|
| |
|
| Submission date |
Jan 21, 2016 |
| Last update date |
Aug 01, 2016 |
| Contact name |
Ryo Inoue |
| E-mail(s) |
r-inoue@kpu.ac.jp
|
| Organization name |
Kyoto Prefectural University
|
| Street address |
1-5, Hangi-cho, shimogamo
|
| City |
Kyoto |
| ZIP/Postal code |
606-8522 |
| Country |
Japan |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE77103 |
Gene expression in peripheral blood mononuclear cells from healthy control and autistic infants |
|
