|
| Status |
Public on Dec 27, 2017 |
| Title |
Test Group 4 |
| Sample type |
RNA |
| |
|
| Source name |
Lgr5-EGFP-IRES-CreER reporter mice, fucosylated, Lgr5+ mouse stem cells
|
| Organism |
Mus musculus |
| Characteristics |
cell type: Lgr5+ mouse stem cells
treatment: fucosylation
|
| Treatment protocol |
Treated organoids were fucosylated daily for 30 mins using H. pylory 1,3 Fucosyltransferase and GDP-Fucose
|
| Growth protocol |
Crypt isolation and organoid preparation were prepared using standard protocols. Organoids were maintained in crypt culture medium (Advanced DMEM/F12, 2 mM GlutaMax, 10 mM Hepes, 1X PenStrep, 1X N2, 1X B27, 1 uM N-acetylcysteine, 50ng/ml EGF, 100 ng/ml Noggin, 1 ug/ml R-spondin). EGF, Noggin and R- spondin were replenished every 2 days, and the full media was changed every 4 days.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated using QIAShredder columns and a RNA Ready Micro kit (Qiagen) according to manufacturer's instructions
|
| Label |
SYBR Green
|
| Label protocol |
RNA was reverse transcribed and amplified with a RT2 PreAMP kit (Qiagen)
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Fucosylated
M4
|
| Data processing |
CT values were exported to an Excel file to create a table of CT values. This table was then uploaded on to the data analysis web portal at http://www.qiagen.com/geneglobe. Samples were assigned to controls and test groups. CT values were normalized with an arithmetic mean of houskeeping genes B2m, Gapdh and Hsp90ab1 included in the array.
Geneglobe software does not make normalized values available only final fold-change. Fold-change values for fucosylated/untreated organoids.
|
| |
|
| Submission date |
Dec 26, 2017 |
| Last update date |
Dec 27, 2017 |
| Contact name |
Aime Lopez Aguilar |
| Organization name |
The Scripps Research Institute
|
| Department |
Molecular Medicine
|
| Lab |
Wu Lab
|
| Street address |
10550 North Torrey Pines Road
|
| City |
La Jolla |
| State/province |
CA |
| ZIP/Postal code |
92037 |
| Country |
USA |
| |
|
| Platform ID |
GPL24429 |
| Series (1) |
| GSE108512 |
Engineered glycocalix regulates stem cell proliferation in murine crypt organoids |
|
