|
| Status |
Public on Mar 07, 2014 |
| Title |
99WT GERM CELL PELLET |
| Sample type |
RNA |
| |
|
| Source name |
Wildtype germ cells
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
mouse age: 5 months
proband sex: Male
tissue: cell isolation from testis
|
| Treatment protocol |
mice were euthanized with carbon dioxide and the whole testis was removed. Germ cells and Sertoli cells were isolated.
|
| Growth protocol |
animals were housed at the mouse facility of the ENI, University Göttingen
|
| Extracted molecule |
total RNA |
| Extraction protocol |
mRNA was isolated from the germ and Sertoli cell population, which were isolated from murine WT and TAp73KO testis. Cell samples were transferred into trizol buffer, homogenized and mRNA was isolated in a single-step acid guanidinium thiocyanate-phenol-chloroform extraction following manufacturer´s instructions (http://tools.invitrogen.com/content/sfs/manuals/trizol_reagent.pdf). RNA-precipitation was done with Ethanol. RNA was diluted in nuclease-free water and concentrations were determined using a Nanodrop spectrophotometer.
|
| Label |
biotin
|
| Label protocol |
For each array 200 ng RNA was processed and Cy3 labeled by the Low Input Quick Amp Labeling Kit (Agilent Technologies).
|
| |
|
| Hybridization protocol |
Hybridization was performed in the hybridization oven (Agilent Technologies, 17h, 10 rpm, 65˚C)
|
| Scan protocol |
Cy3 intensities were detected by a one-color microarray scanner (G2505C, Agilent Technologies) at 5 µm resolution
|
| Data processing |
Data were quantile normalized and log2 transformed using R and Bioconductor package limma.
|
| |
|
| Submission date |
Nov 06, 2013 |
| Last update date |
Mar 07, 2014 |
| Contact name |
Gabriela Salinas |
| E-mail(s) |
[email protected]
|
| Organization name |
Universitaetsmedizin Goettingen
|
| Department |
Department of Pathology
|
| Lab |
NGS Integrative Genomics
|
| Street address |
Kreuzbergring 57
|
| City |
Goettingen |
| State/province |
Lower-Saxony |
| ZIP/Postal code |
37075 |
| Country |
Germany |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE52124 |
TAp73 is essential for germ cell adhesion and maturation in testis |
|
