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Status |
Public on Jan 11, 2013 |
Title |
In vitro-differentiated human Th2 cells |
Sample type |
RNA |
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Source name |
Human Th2 polarized cells
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Organism |
Homo sapiens |
Characteristics |
cell type: in vitro Th2 cells (derived from blood CD4 T-cells)
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Treatment protocol |
Blood derived primary CD4+ T-cells were differentiated in vitro into a TH2 phenotype (Sandig et al. 2007. Human th2 cells selectively express the orexigenic peptide, pro-melanin-concentrating hormone. Proc Natl Acad Sci U S A 104: 12440-12444)
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Extracted molecule |
total RNA |
Extraction protocol |
The total RNA extraction was performed using the miRNeasy kit from Qiagen (including theoptional on column-DNAseI treatment). RNA quantity was measure by OD spectrophotometry (Nanodrop).
|
Label |
Cy3
|
Label protocol |
Total RNA was labeled with Cy3-CTP using the miRCURY LNA microRNA power labeling kit (Exiqon, Inc, Woburn, MA), according to manufacturers protocol
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Hybridization protocol |
Cy3-labelled RNA was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent UCSF custom 8x15K miRNA arrays for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
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Scan protocol |
Slides were scanned on the Agilent scanner immediately after washing.
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Description |
Raw data file: 252372310006_S01_miRNA-v1_95_May07_2_2.txt
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Data processing |
The scanned images were analyzed with Feature Extraction Software 10.6 (Agilent) using default parameters) All replicates of human miR probes have been collapsed and median signal intensity extracted.
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Submission date |
Jul 29, 2011 |
Last update date |
Jan 11, 2013 |
Contact name |
Andrea J Barczak |
Organization name |
UCSF Sandler Center Functional Genomics Core Facility
|
Street address |
1550 4th St Rock Hall 545
|
City |
San Francisco |
State/province |
CA |
ZIP/Postal code |
94143 |
Country |
USA |
|
|
Platform ID |
GPL13992 |
Series (1) |
GSE31030 |
Targeting microRNAs for asthma treatment |
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