|
| Status |
Public on Mar 27, 2019 |
| Title |
uterosacral ligament_primary pelvic organ prolapse_subject 4 |
| Sample type |
RNA |
| |
|
| Source name |
uterosacral ligament_primary POP_subject 4
|
| Organism |
Homo sapiens |
| Characteristics |
Stage: postmenopausal
gender: female
tissue: uterosacral ligament (USL)
treatment: none
pathology: primary pelvic organ prolapse (POP)
popq stage: stage III
subject identifier: primary POP 4
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Uterosacral ligament (USL) tissue samples were obtained at surgery. The tissue samples were placed in RNAlater (Ambion) in the surgical suite. For processing, RNAlater was blotted off and the USL sample was minced with scalpels, and soaked in TRI reagent for 1 hour. The sample was homogenized in TRI reagent (MRC) with a Polytron (Brinkmann). The samples were centrifuged in the presence of bromochloropropane and sodium acetate. The RNA-containing upper layer was removed and the RNA purified on an RNeasy extraction column (Qiagen). The sample was treated with an on-column DNase treatment (RNase-free DNase, Qiagen). The purity and quantity of total RNA was evaluated with the RNA 6000 NanoLabChip in an Agilent Bioanalyzer.
|
| Label |
Alexa-647
|
| Label protocol |
Biotinylated cRNA was prepared using the MessageAmp II-Biotin Enhanced Kit from Ambion from 0.75 micrograms total RNA starting material.
|
| |
|
| Hybridization protocol |
10 micrograms of biotinylated cRNA were mixed with fragmentation buffer and heated to 94C for 20 min. The fragmented cRNA was mixed with CodeLink hybridization buffer, loaded on the microarray slides, and hybridized for 18 hours at 37C.
|
| Scan protocol |
The slides were scanned in an Axon GenePix 4000B scanner.
|
| Description |
species: human subject, stage:postmenopausal, gender:female, tissue:uterosacral ligament biopsy, treatment:none, pathology:primary pelvic organ prolapse, POPQ stage:stage III, sample:primary POP 4
Approval for the use of human subjects was obtained from the Institutional Review Board and patients provided written informed consent.
|
| Data processing |
global median normalization, CodeLink Expression Analysis 5.0
|
| |
|
| Submission date |
Apr 14, 2011 |
| Last update date |
Mar 27, 2019 |
| Contact name |
Kathleen M Eyster |
| E-mail(s) |
Kathleen.Eyster@usd.edu
|
| Organization name |
University of South Dakota
|
| Department |
Basic Biomedical Sciences
|
| Street address |
414 E. Clark St.
|
| City |
Vermillion |
| State/province |
SD |
| ZIP/Postal code |
57069 |
| Country |
USA |
| |
|
| Platform ID |
GPL2895 |
| Series (1) |
| GSE28660 |
Differential gene expression in uterosacral ligament from patients with recurrent and primary pelvic organ prolapse |
|
