|
| Status |
Public on May 05, 2017 |
| Title |
Subpopulation P5 (Lin-CD24+CD29lo) from virgin wild type FVB mice, replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
Luminal enriched P5 population of wild type virgin mice
|
| Organism |
Mus musculus |
| Characteristics |
strain: FVB/N-J
mg condition: Virgin
enrichment: Luminal progenitor enriched P5 population
genotype/variation: wild type
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cell preparation protocol: Single cell suspensions were prepared from the mammary glands of virgin wild type and transgenic mice using mechanical and enzymatic dissociation as detail described in Tiede BJ et al., 2009, PLoS ONE 4(11): e8035. doi:10.1371/journal.pone.0008035
FACS sorting protocol: Various subpopulations of mammary epithelial cells were sorted using flow cytometry based on combination of a set of surface markers. The marker profiles and FACS gating parameters are detailed in Tiede BJ et al., 2009, PLoS ONE 4(11): e8035. doi:10.1371/journal.pone.0008035
miRNA enriched Total RNA were extracted using mirVana™ miRNA Isolation Kit from Ambion
|
| Label |
Cy3
|
| Label protocol |
The testing RNA samples were labeled with Cy3-pCp using Agilent’s miRNA Complete Labeling and Hyb Kit. Briefly, total RNA is dephosphrylated with CIP and then ligated to Cy3-pCp with T4-RNA ligase. Labeled RNA sample were purified and dried.
|
| |
|
| Hybridization protocol |
Using Agilent’s miRNA Complete Labeling and Hyb Kit, labeled total RNA samples were mixed with blocking reagent and RPM hybridization buffer and assembled with miRNA array for hybridization. The hybridization assembly were rotate at 20rpm in 55C oven for 20hrs. The array were washed with prewarmed washing buffers and dried for scan.
|
| Scan protocol |
Scanned on an Agilent G2505C scanner.
Images were quantified using Agilent Feature Extraction Software (version 11.0).
|
| Data processing |
Agilent Feature Extraction Software (v 11.0) was used to extract the signal and GeneSpring v13 were used to process the data. The processing is using Agilent technology miRNA 46065 with Threshold 1.0, logbase 2, Normalization shift to 90 percentile and baseline transformation is conducted based on median of all samples.
|
| |
|
| Submission date |
Aug 15, 2016 |
| Last update date |
Feb 29, 2024 |
| Contact name |
Yong Wei |
| E-mail(s) |
[email protected]
|
| Organization name |
Princeton University
|
| Department |
Molecular Biology
|
| Lab |
Kang
|
| Street address |
LTL227 Wathington Road
|
| City |
Plainsboro |
| State/province |
NJ |
| ZIP/Postal code |
08536 |
| Country |
USA |
| |
|
| Platform ID |
GPL17912 |
| Series (2) |
| GSE85634 |
microRNA profiles of mouse mammary stem cells and luminal cells |
| GSE85808 |
miR-199a-LCOR axis regulation of mammary gland stem cell and breast cancer stem cell transcriptional programs |
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