|
| Status |
Public on Jan 05, 2011 |
| Title |
Kc167Cells_HSFIP_2nd_half_rep1 |
| Sample type |
genomic |
| |
|
| Channel 1 |
| Source name |
anti-HSF
|
| Organism |
Drosophila melanogaster |
| Characteristics |
cell type: Kc167 cell line
antibody: anti-HSF
|
| Treatment protocol |
ip: chromosal DNA was obtained by chromatin-imunoprecipitation as described by Weinmann and Farnham 2002 with an anti-HSF antibody
mock: chromosal DNA was obtained by mock chromatin-imunoprecipitation as described by Weinmann and Farnham 2002 with no antibody
WCE: chromosal DNA was obtained as described by Weinmann and Farnham 2002 with an anti-HSF antibody
|
| Growth protocol |
Kc167 cells were grown at 22C in Schneider's medium supplemented with 5% heat-inactivated FBS and 20ug/ml gentamicin in tissue culture flasks to a final concentration of ~1x10^7 cells per ml.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
According to Agilent protocol
|
| Label |
Alexa 647
|
| Label protocol |
Anti-HSF or mock immunoprecipitated chromosomal DNA was subjected to two rounds of amplification by LM-PCR as described by Ren et al. 2000 and Birch-Machin et al. 2005 with amino-allyl modified nucleotides being incorporated into the amplified product in the second round. Fluorescent dye conjugation and probe clean-up and precipitation was performed as described on the CDMC web site (www.flyarrays.com).
|
| |
|
| Channel 2 |
| Source name |
Whole Cell Extract (WCE)
|
| Organism |
Drosophila melanogaster |
| Characteristics |
cell type: Kc167 cell line
antibody: none (whole cell extract)
|
| Treatment protocol |
ip: chromosal DNA was obtained by chromatin-imunoprecipitation as described by Weinmann and Farnham 2002 with an anti-HSF antibody
mock: chromosal DNA was obtained by mock chromatin-imunoprecipitation as described by Weinmann and Farnham 2002 with no antibody
WCE: chromosal DNA was obtained as described by Weinmann and Farnham 2002 with an anti-HSF antibody
|
| Growth protocol |
Kc167 cells were grown at 22C in Schneider's medium supplemented with 5% heat-inactivated FBS and 20ug/ml gentamicin in tissue culture flasks to a final concentration of ~1x10^7 cells per ml.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
According to Agilent protocol
|
| Label |
Alexa 555
|
| Label protocol |
Anti-HSF or mock immunoprecipitated chromosomal DNA was subjected to two rounds of amplification by LM-PCR as described by Ren et al. 2000 and Birch-Machin et al. 2005 with amino-allyl modified nucleotides being incorporated into the amplified product in the second round. Fluorescent dye conjugation and probe clean-up and precipitation was performed as described on the CDMC web site (www.flyarrays.com).
|
| |
|
| |
| Hybridization protocol |
According to Agilent protocol
|
| Scan protocol |
Images were quantified using Agilent Feature Extraction Software (version A.8.5.1.1) according to Agilent protocol.
|
| Description |
Anti-HSF immunoprecipitated chromosomal DNA
|
| Data processing |
Agilent Feature Extraction Software (v 8.5.1.1) was used for background subtraction, Blank subtraction normalization, Inter-array median normalization, Intra-array (dye-bias) median normalization
|
| |
|
| Submission date |
Jan 05, 2010 |
| Last update date |
Jan 05, 2011 |
| Contact name |
Zak Razak |
| E-mail(s) |
z.razak@utoronto.ca
|
| Phone |
905-569-4664
|
| Fax |
905-828-3792
|
| URL |
http://www.flyarrays.com
|
| Organization name |
Canadian Drosophila Microarray Centre
|
| Street address |
3359 Mississauga Rd
|
| City |
Mississauga |
| State/province |
Ontario |
| ZIP/Postal code |
L5L 1C6 |
| Country |
Canada |
| |
|
| Platform ID |
GPL4150 |
| Series (2) |
| GSE19744 |
Whole-genome analysis of heat shock factor binding sites in Drosophila melanogaster during heat shock |
| GSE19746 |
Whole-genome analysis reveals active heat shock factor binding sites are mostly associated with non-heat shock genes |
|
