|
| Status |
Public on May 09, 2014 |
| Title |
epithelial_116-120h_rep2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
pooled reference amplified RNA from whole animals and sorted populations
|
| Organism |
Drosophila melanogaster |
| Characteristics |
tissue: Whole animal
|
| Growth protocol |
Timed larvae collections on grape juice agar plates
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Dissociated cells isolated by FACS. Up to 10k cells sorted into Ambion RNAqueous Micro kit lysis buffer. RNA isolated as per manufacturer's instructions with DNase treatment. RNA amplified in two rounds with Ambion Aminoallyl MessageAmp II kit.
|
| Label |
Cy3
|
| Label protocol |
1.5 ug of aRNA coupled to Cy5 (or Cy3 for common pooled reference aRNA) as per standard protocols
|
| |
|
| Channel 2 |
| Source name |
Amplified RNA from FACS-sorted epithelial cells 116-120h AEL
|
| Organism |
Drosophila melanogaster |
| Characteristics |
cell type: Epithelia
marker: arm
age (hours after egg laying): 116-120h
|
| Growth protocol |
Timed larvae collections on grape juice agar plates
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Dissociated cells isolated by FACS. Up to 10k cells sorted into Ambion RNAqueous Micro kit lysis buffer. RNA isolated as per manufacturer's instructions with DNase treatment. RNA amplified in two rounds with Ambion Aminoallyl MessageAmp II kit.
|
| Label |
Cy5
|
| Label protocol |
1.5 ug of aRNA coupled to Cy5 (or Cy3 for common pooled reference aRNA) as per standard protocols
|
| |
|
| |
| Hybridization protocol |
825 ng of each aRNA hybridized to custom Agilent 4x44k microarrays as per manufacturer's instructions
|
| Scan protocol |
Molecular Devices Genepix 4000B scanner at 5 um resolution
|
| Data processing |
Software: GenePix 6.0. Ratio normalized, filters: 'B532' <= 350 AND 'B635' <= 350) AND 'F532 % Sat.' <= 60 AND 'F635 % Sat.' <= 60 AND 'F532 CV' <= 80 AND 'F635 CV' <= 80 AND 'Flags' >= 0 AND 'Dia.' >= 50 AND 'Dia.' <= 80 AND ('% > B532+2SD' >= 70 OR '% > B635+2SD' >= 70) AND 'Substance Name' begins with 'f' AND 'Circularity' >= 60
|
| |
|
| Submission date |
Jul 30, 2010 |
| Last update date |
May 09, 2014 |
| Contact name |
Charlie Kim |
| E-mail(s) |
cckim47@gmail.com
|
| Organization name |
University of California, San Francisco
|
| Department |
Medicine
|
| Lab |
Kim
|
| Street address |
1001 Potrero Ave
|
| City |
San Francisco |
| State/province |
CA |
| ZIP/Postal code |
94110 |
| Country |
USA |
| |
|
| Platform ID |
GPL10744 |
| Series (1) |
| GSE23320 |
Gene Expression Dynamics of Defined Cell Populations in Developing Drosophila Embryos and Larvae |
|
| Relations |
| Reanalyzed by |
GSM1220911 |
