|
| Status |
Public on Dec 05, 2013 |
| Title |
Gemin2 MO replicate 5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Gemin2 MO injected
|
| Organism |
Danio rerio |
| Characteristics |
tissue: whole embryo
Stage: 48hpf
|
| Treatment protocol |
Embryos were injected with either SMN MO, Gemin2 MO, or control MO at one - two cell stage
|
| Growth protocol |
Zebrafish embryos reared at temperature of 28°C in embryo media
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted after homogenisation and Phenol Chlorofoam extraction, DNase I digestion followed by second round of Phenol-Chlorofoam extraction
|
| Label |
Cy5
|
| Label protocol |
To fluorescently label cDNA, 10μg of total RNA per experimental sample and per control sample was reverse transcribed in the presence of Cy5 and Cy3 dye (Amersham Inc.) respectively. Labelled cDNAs were pooled, concentrated and purified using QIAquick columns before being resuspended in DIG EasyHyb (Roche) for hybridisation.
|
| |
|
| Channel 2 |
| Source name |
Control MO injected
|
| Organism |
Danio rerio |
| Characteristics |
tissue: whole embryo
Stage: 48hpf
|
| Treatment protocol |
Embryos were injected with either SMN MO, Gemin2 MO, or control MO at one - two cell stage
|
| Growth protocol |
Zebrafish embryos reared at temperature of 28°C in embryo media
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted after homogenisation and Phenol Chlorofoam extraction, DNase I digestion followed by second round of Phenol-Chlorofoam extraction
|
| Label |
Cy3
|
| Label protocol |
To fluorescently label cDNA, 10μg of total RNA per experimental sample and per control sample was reverse transcribed in the presence of Cy5 and Cy3 dye (Amersham Inc.) respectively. Labelled cDNAs were pooled, concentrated and purified using QIAquick columns before being resuspended in DIG EasyHyb (Roche) for hybridisation.
|
| |
|
| |
| Hybridization protocol |
Hybridisation was performed at 42°C overnight in MAUI hybridisation station (BioMicro)
|
| Scan protocol |
Scanning of the arrays was performed using GenePix Pro 4.0 image analysis software on a GenePix 4000B microarray scanner (Axon Instruments, California, US).
|
| Description |
Biological replicate 5 of 7
|
| Data processing |
Genespring v10 software was used to generate LOWESS normalized data from log ratio of processed Cy5 signal/processed Cy3 signal.
|
| |
|
| Submission date |
May 16, 2013 |
| Last update date |
Dec 05, 2013 |
| Contact name |
Kelvin See |
| Organization name |
University of Pennsylvania
|
| Department |
Medicine
|
| Street address |
3400 Civic Center Blvd, Smilow Center for Translational Research
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL10182 |
| Series (1) |
| GSE47001 |
SMN MO, Gemin2 MO knockdown vs Control MO injected |
|
