|
| Status |
Public on May 03, 2012 |
| Title |
X153L.p11AM87ACTX_Pre-stasis HMEC Biological replicate #1 |
| Sample type |
RNA |
| |
|
| Source name |
pre-stasis HMEC at stasis
|
| Organism |
Homo sapiens |
| Characteristics |
age: 60
tissue source: human mammary gland (reduction mammoplasty tissue)
cell type: pre-stasis HMEC at stasis
passage number: 11
±oxytocin: +oxytocin
|
| Treatment protocol |
Cell cultures were harvested and different points in their growth from early passage good growing to near stasis.
|
| Growth protocol |
Finite life span HMEC were obtained from reduction mammoplasty tissue of women of the indicated ages. HMEC were grown starting from organoids in the serum-containing M85+CT±X or M87A+CT+X medium.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cell were lysed with trizol reagent (Invitrogen) according manufacturer's protocol. RNA was resuspended in 100 µl nuclease free water and subject to a second step purification on ion exchange column (Qiagen).
|
| Label |
biotin
|
| Label protocol |
For each sample, the RNA target is prepared by annealing 2.0µg of total RNA in 5µl water with 5µl of 10µM T7 (Oligo dt) primer. First and second strand cDNA synthesis is performed and the cDNA products purified using magnetic beads. Labeled cRNA is synthesized from the cDNA using T7 RNA polymerase followed by clean-up with magnetic beads. Purified cRNA is quantitated and fragmented.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip HT_HG-U133A Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Hewlett-Packard GeneArray Scanner G2500A.
|
| Description |
X153L.p11AM87ACTX
|
| Data processing |
Robust multi-array analysis (RMA, from affy package version 1.26.1) was performed to normalize data collected from different samples through R bioconductor (R version 2.11.1). Default parameter values were used.
|
| |
|
| Submission date |
Apr 24, 2012 |
| Last update date |
May 04, 2012 |
| Contact name |
Francois Pepin |
| Organization name |
McGill University
|
| Street address |
1160 Pine Av W
|
| City |
Montreal |
| State/province |
Quebec |
| ZIP/Postal code |
H3A1A3 |
| Country |
Canada |
| |
|
| Platform ID |
GPL3921 |
| Series (1) |
| GSE37485 |
Expression data of pre-stasis cultured HMEC |
|
