|
| Status |
Public on Apr 01, 2015 |
| Title |
mir-3 ectopic replicate 2 vs. w1118 replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
mir-3 ectopic expressed embryo
|
| Organism |
Drosophila melanogaster |
| Characteristics |
Stage: 6-8hr after egg laying
genetic background: w1118
genotype: miR-3 ectopic expressing mutant
tissue: embryo
|
| Treatment protocol |
wild type and mir-3 ectopic expressed embryos were treated same.mutant embryos are homozygous for mir-3 ectopic expression.
|
| Growth protocol |
Wild type (w1118) and mir-3 ectopic expressed mutant flies were maintained at 25oC and collected embryo for 2 hours. After egg laying the embryo were kept at 6 hours at 25oC.Then isolated total RNA.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Total RNA was extracted with Trizol reagent (Invitrogen) according to the manufacturer's instructions and then passed through the RNeasy Mini Kit columns(Qiagen) after DNAse I digestion. RNA quality was monitored using Agilent RNA Bioanalyzer 2100 analysis and RNA quantification was carried out on a Nanodrop ND-1000. Array analysis was performed according to the manufacturers instructions using the Quick Amp Labeling kit (Agilent).
|
| |
|
| Channel 2 |
| Source name |
w1118 embryo
|
| Organism |
Drosophila melanogaster |
| Characteristics |
Stage: 6-8hr after egg laying
genetic background: w1118
genotype: wild type
tissue: embryo
|
| Treatment protocol |
wild type and mir-3 ectopic expressed embryos were treated same.mutant embryos are homozygous for mir-3 ectopic expression.
|
| Growth protocol |
Wild type (w1118) and mir-3 ectopic expressed mutant flies were maintained at 25oC and collected embryo for 2 hours. After egg laying the embryo were kept at 6 hours at 25oC.Then isolated total RNA.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
Total RNA was extracted with Trizol reagent (Invitrogen) according to the manufacturer's instructions and then passed through the RNeasy Mini Kit columns(Qiagen) after DNAse I digestion. RNA quality was monitored using Agilent RNA Bioanalyzer 2100 analysis and RNA quantification was carried out on a Nanodrop ND-1000. Array analysis was performed according to the manufacturers instructions using the Quick Amp Labeling kit (Agilent).
|
| |
|
| |
| Hybridization protocol |
Quick Amp Labeling kit (Agilent) Total RNA samples were labeled with Cy3 and Cy5, respectively, and hybridized to Agilent Drosophila melanogaster 4-by-44000 60-mer-oligonucleotide array slides and enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed and scanned.
|
| Scan protocol |
Scanned on an Agilent G2565AA scanner.
|
| Description |
First biological replicate is dye swaped to avoid dye bias
|
| Data processing |
Images were quantified using Agilent Feature Extraction Software (version A.7.5). The data was then processed with Gene Spring data analysis program.
|
| |
|
| Submission date |
Apr 13, 2011 |
| Last update date |
Apr 01, 2015 |
| Contact name |
Anandarao Ravulapalli |
| E-mail(s) |
ravulapa@mail.nih.gov
|
| Phone |
301-594-3314
|
| Organization name |
NCI
|
| Department |
LBMB
|
| Street address |
9000 Rockville Pike
|
| City |
Bethesda |
| State/province |
maryland |
| ZIP/Postal code |
20852 |
| Country |
USA |
| |
|
| Platform ID |
GPL7300 |
| Series (2) |
| GSE28605 |
Ectopic expression of mir-3 affects mef2 and tinman levels |
| GSE28614 |
Nautilus and muscle development |
|
