|
| Status |
Public on Jul 17, 2016 |
| Title |
Blood CD14+ monocyte, rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Blood CD14+ monocytes
|
| Organism |
Homo sapiens |
| Characteristics |
tissue source: blood
cell type: CD14+ monocytes
sample type: pooled
|
| Treatment protocol |
Blood CD14+ monocytes were isolated from peripheral blood mononuclear cells of healthy volunteers using flow cytometry.
|
| Growth protocol |
None
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cells using an miRNeasy Mini kit according to the manufacturer’s instructions (Qiagen). The quality of the RNAs was estimated by using the RNA 6000 Nano LabChip Kit on the Agilent 2100 Bioanalyzer (Agilent).
|
| Label |
Cy3
|
| Label protocol |
50 ng of total RNAs were independently reverse-transcribed using oligo-dT primers containing the T7 RNA polymerase promoter sequence to generate cDNAs and AffinityScript, RTase, which were then subjected to in vitro transcription using T7 RNA polymerase to label the cRNAs with Cy3-CTP (Amersham Pharmacia Biotech, Piscataway, NJ) using a Low input Quick-Amp Labeling Kit (Agilent Technologies).
|
| |
|
| Hybridization protocol |
Before hybridization, 825 ng labeled cRNA of each product was fragmented and mixed with control targets and hybridization buffer according to the manufacturer's protocol (Agilent Technologies). Hybridizations were done for approximately 17h at 65°C. The slides were washed according to the manufacturer's manual.
|
| Scan protocol |
The array was scanned with 5µm resolution using an Agilent G2505C DNA microarray scanner. Images were quantified using Agilent Feature Extraction Software (version 10.5.1.1).
|
| Description |
Monocyte_2
|
| Data processing |
Data were analyzed using the bioinformatics software GeneSpring 13.0 (Agilent).
|
| |
|
| Submission date |
Mar 25, 2016 |
| Last update date |
Jul 17, 2016 |
| Contact name |
Daisuke Okuzaki |
| E-mail(s) |
dokuzaki@biken.osaka-u.ac.jp
|
| Phone |
+81-6-6879-4935
|
| Organization name |
Osaka univ.
|
| Department |
Immunology Frontier Research Center
|
| Lab |
Human Immunology （Single Cell Genomics)
|
| Street address |
Yamadaoka 3-1
|
| City |
Suita |
| State/province |
Osaka |
| ZIP/Postal code |
565-0871 |
| Country |
Japan |
| |
|
| Platform ID |
GPL4133 |
| Series (2) |
| GSE79627 |
Microarray analysis of CD14+ CD163high CD160high cells in human intestinal lamina propria |
| GSE79629 |
Microarray analysis of CD14+ CD163high CD160high cells in human intestinal lamina propria and CX3CR1high CD11b+ CD11c+ cells in murine intestinal lamina propria |
|
