|
| Status |
Public on Apr 27, 2007 |
| Title |
Teratozoospermic individual: Sample 23D |
| Sample type |
RNA |
| |
|
| Source name |
Human Sperm from an infertile male with a consistent severe teratozoospermia
|
| Organism |
Homo sapiens |
| Characteristics |
Fertility Status: Infertile
|
| Treatment protocol |
None
|
| Growth protocol |
Human donor
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Semen was collected then treated with Somatic Cell Lysis Buffer (0.1% sodium dodecyl sulfate [SDS], 0.5% Triton X-100) on ice for 30 minutes. Lack of somatic cell contamination and sperm count were established by microscopic examination. Samples were lysed and RNAs stabilized in a chaotropic buffer. RNA was isolated from sample using a modified RNeasy protocol (Qiagen Inc. Valencia, CA). RNA was eluted from collection columns using 50 microL of nuclease-free water twice for a final volume of 100 microL. Dithiothreitol (DTT) was added to eluted samples to a final concentration of 10 mM. A total of 40 units RNase block (Strategene, La Jolla, CA) was added to each sample prior to a 20-minute treatment with RNase-free DNase I (Ambion, Austin, TX) at 37°C.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Illumina protocol from total RNA (Whole-Genome Gene Expression with IntelliHyb Seal System Manual, 2006, Illumina).
|
| |
|
| Hybridization protocol |
Hybridization to Illumina Sentrix Expression BeadChip, staining and washing were carried out as described (Illumina, Wallingford).
|
| Scan protocol |
Scanning procedures were carried out as described by Illumina, Wallingford.
|
| Description |
Spermatozoal RNA profile from an infertile individual with a consistent severely teratozoospermic semen profile.
|
| Data processing |
The data were analyzed with Illumina Beat Studio using Cubic Spline Normalization
|
| |
|
| Submission date |
Feb 06, 2007 |
| Last update date |
Apr 27, 2007 |
| Contact name |
Stephen A Krawetz |
| E-mail(s) |
steve@compbio.med.wayne.edu
|
| Organization name |
Wayne State University
|
| Street address |
275 East Hancock, CS Mott Center
|
| City |
Detroit |
| State/province |
MI |
| ZIP/Postal code |
48207 |
| Country |
USA |
| |
|
| Platform ID |
GPL2700 |
| Series (2) |
| GSE6968 |
Spermatozoal RNA Profiles (HumanRef-8 BeadChip) |
| GSE6969 |
Spermatozoal RNA Profiles from Normally Fertile and Heterogeneously Teratozoospermic Subjects |
|
