|
| Status |
Public on Sep 24, 2007 |
| Title |
Wild type embryos rep2 |
| Sample type |
RNA |
| |
|
| Source name |
WT embryos 3-5 hrs AEL. Repetition2.
|
| Organism |
Drosophila melanogaster |
| Characteristics |
yw
|
| Treatment protocol |
. Homozygous how mutant embryos were separated from their siblings using a fluorescent microscope. Carefully staged embryos which had been aged 3–5 hr in this manner were collected and dechorionated.
|
| Growth protocol |
Two hour embryo collections were made from either YW or from how germline clone mutant females on apple juice/yeast plates at 25°C. Plates were removed and the embryos were aged an additional 3 hr at 25°C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from sufficient amounts of embryos ( 100 embryos) using the Macherey-Nagel's:Nucleospin RNA II Mini Kit following the protocol, and then kept in -70°C. Total RNA was prepared independently five times from embryos of each genetic background in order to better normalize the age of these embryo populations. The RNA samples were then collected and concentrated to give 1mg of total RNA using the RNA cleanup RNeasy Mini Kit (Qiagen).
|
| Label |
biotin
|
| Label protocol |
The probe Preparation, cDNA synthesis, cRNA reactions and hybridization with Affymetrix high-density oligonucleotide arrays for Drosophila melanogaster was carried out in the Weizmann Institute microarray unit.
|
| |
|
| Hybridization protocol |
Following fragmentation, 10 ug of cRNA were hybridized for 16 hr at 45C on GeneChip Drosophila Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned using Affymetrix 3000G.
|
| Description |
Gene expression data from yw embryos 3-5 h AEL.
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was arbitarily set to 250.
|
| |
|
| Submission date |
May 10, 2007 |
| Last update date |
Sep 24, 2007 |
| Contact name |
Talila Volk |
| E-mail(s) |
[email protected]
|
| Phone |
972-8-9342426
|
| Fax |
972-8-9344108
|
| Organization name |
Weizmann Institute
|
| Department |
Molecular Genetics
|
| Street address |
Hertzel street
|
| City |
Rehovot |
| ZIP/Postal code |
76100 |
| Country |
Israel |
| |
|
| Platform ID |
GPL72 |
| Series (1) |
| GSE7772 |
Comparison between mRNAs of how germ-line clones embryos and WT embryos at 3-5 h AEL |
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