|
| Status |
Public on Sep 30, 2015 |
| Title |
Neg CD5- CD43-_1 |
| Sample type |
RNA |
| |
|
| Source name |
CD5 negative B lymphoyctes
|
| Organism |
Homo sapiens |
| Characteristics |
phenotype: CD20+CD19+CD3-CD5-CD10-
cell type: CD5 negative B lymphoyctes
|
| Treatment protocol |
Mononuclear cells were isolated by Ficoll from heparinized cord blood, and then sorted by FACS to purify B lymphocytes, and cell pellet directly snap frozen
|
| Growth protocol |
freshly isolated cells, no culture
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was extracted using the NucleoSpin® RNA II kit (Macherey & Nagel) following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
10 ng of each RNA was used as template to produce Cy3- labeled cRNA. The RNA samples were amplified and labeled using the Low Input Quick Amp Labeling Kit (Agilent Technologies) following the manufacturer’s protocol.
|
| |
|
| Hybridization protocol |
Cy3 labeled cRNAs were hybridized overnight (17 hours, 65°C) to Agilent Whole Human Genome Oligo Microarrays 8x60K using Agilent’s recommended hybridization chamber and oven.Finally, microarrays were washed once with wash buffer 1 for 1 min at RT followed by a second wash with wash buffer 2 for 1 min at 37°C and a final wash step with acetonitrile for 30 sec at RT.
|
| Scan protocol |
Fluorescence signals of the hybridized Agilent Microarrays were detected using Agilent’s Microarray Scanner System (G2505C, Agilent Technologies, Palo Alto, USA).
|
| Description |
cord blood 262
|
| Data processing |
The Agilent Feature Extraction Software (FES 10.7.3.1 ) was used to read out and process the microarray image files.
|
| |
|
| Submission date |
Jul 19, 2012 |
| Last update date |
Sep 30, 2015 |
| Contact name |
Silvia Rueberg |
| E-mail(s) |
silvia@miltenyibiotec.de
|
| Organization name |
Miltenyi Biotec GmbH
|
| Department |
Genomic Services
|
| Street address |
Friedrich-Ebert-Str. 68
|
| City |
Bergisch Gladbach |
| ZIP/Postal code |
51429 |
| Country |
Germany |
| |
|
| Platform ID |
GPL14550 |
| Series (1) |
| GSE39489 |
Analysis of neonatal B lymphocytes |
|
