|
| Status |
Public on Mar 30, 2016 |
| Title |
euploid_pre_mid_pach_SC_1 |
| Sample type |
RNA |
| |
|
| Source name |
t121/D17_pre-mid-pachytene spermatocytes
|
| Organism |
Mus musculus |
| Characteristics |
genotype: t121/D17
gender: male
age: adult (2 moths)
tissue: pre-mid-pachytene spermatocytes
|
| Treatment protocol |
Mice were euthanized by cervical dislocation. For isolation of spermatogenic populations the single cell suspension from testicular tubules was prepared and the populations were isolated using fluorescence activated cell sorting of testicular single cell suspension according to ref. (Bastos et al. 2005).
|
| Growth protocol |
All the male mice used for isolation of spermatogenic populations were two months old.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using RNeasy Micro kit (Qiagen) according to manufacturer‘s protocol.
|
| Label |
biotin
|
| Label protocol |
Between 20 to 30 ng of total RNA were used to prepare biotinylated cRNA according to the standard Affymetrix Two-Cycle protocol (Expression Analysis Technical Manual, 701021 Rev. 5, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of cRNA were hybridized for 16 hr at 45°C on GeneChip Mouse Genome 430 2.0 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using the GeneChip Scanner 3000 7G.
|
| Data processing |
The data were analyzed with Bioconductor and R statistics using the gcRMA method with default settings and custom chip description file (probes were annotated to Entrez identifiers - NCBI build 37) mm430mmentrezg, created by Microarray Lab at the Molecular and Behavioral Neuroscience Institute at the University of Michigan.
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| |
|
| Submission date |
Feb 05, 2014 |
| Last update date |
Mar 30, 2016 |
| Contact name |
David Homolka |
| E-mail(s) |
david.homolka@post.cz
|
| Organization name |
Institute of Molecular Genetics AS CR
|
| Street address |
Videnska 1083
|
| City |
Prague |
| ZIP/Postal code |
14220 |
| Country |
Czech Republic |
| |
|
| Platform ID |
GPL7546 |
| Series (1) |
| GSE54683 |
Dosage Compensation of an Aneuploid Genome in Mouse Spermatogenic Cells |
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