|
Status |
Public on Feb 01, 2015 |
Title |
Mutant Huntingtin sorted cells replicate-1 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
128Q embryonic purified touch-receptor cells
|
Organism |
Caenorhabditis elegans |
Characteristics |
cell type: embryonic touch-receptor cells polyq: mutant Huntingtin (128Q)
|
Treatment protocol |
Cells were sorted on a Moflo flow cytometer (Dako) based on GFP fluorescence, recovered in Trizol
|
Growth protocol |
embryonic cells were isolated from gravid adults following lysis in a hypochloride solution. Embryos were dissociated with treatement using chitinase/chymotrypsine then trypsine. Cells were then grown overnight on TESPA-coated glass plate.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA extracted using Trizol LS (Invitrogen) following manufacturer's instructions
|
Label |
Cy5
|
Label protocol |
After a one-round linear amplification with MessageAmp kit (Ambion), RNAs were labelled with the CyScribe Post-labelling kit (Amersham) following manufacturer's instructions.
|
|
|
Channel 2 |
Source name |
19Q embryonic purified touch-receptor cells
|
Organism |
Caenorhabditis elegans |
Characteristics |
cell type: embryonic touch-receptor cells polyq: normal Huntingtin (19Q)
|
Treatment protocol |
Cells were sorted on a Moflo flow cytometer (Dako) based on GFP fluorescence, recovered in Trizol
|
Growth protocol |
embryonic cells were isolated from gravid adults following lysis in a hypochloride solution. Embryos were dissociated with treatement using chitinase/chymotrypsine then trypsine. Cells were then grown overnight on TESPA-coated glass plate.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA extracted using Trizol LS (Invitrogen) following manufacturer's instructions
|
Label |
Cy3
|
Label protocol |
After a one-round linear amplification with MessageAmp kit (Ambion), RNAs were labelled with the CyScribe Post-labelling kit (Amersham) following manufacturer's instructions.
|
|
|
|
Hybridization protocol |
probes were hybridized on dual-color Agilent 22k microarrays following manufacturer's protocol.
|
Scan protocol |
Scanned on an Genepix 4000B scanner (Axon).Images were quantified using GenePix Pro Software.
|
Description |
128Q-19Q-rep1 biological replicate 1 of 3. Embryonic cells purified by cell sorting based on GFP fluorescence.
|
Data processing |
Goulphar was used for background subtraction and LOWESS normalization.
|
|
|
Submission date |
Feb 20, 2012 |
Last update date |
Feb 01, 2015 |
Contact name |
Christian Neri |
E-mail(s) |
christian.neri@inserm.fr
|
Phone |
+33 1 44 27 60 45
|
Organization name |
Sorbonne Université, CNRS, INSERM
|
Department |
CNRS UMR 8256, Inserm ERL U1164
|
Lab |
Brain-C lab
|
Street address |
9 quai St Bernard
|
City |
Paris |
ZIP/Postal code |
75005 |
Country |
France |
|
|
Platform ID |
GPL2875 |
Series (1) |
GSE35939 |
Purified touch receptor neurons: expanded polyGlutamine versus control |
|