|
Status |
Public on Jul 01, 2016 |
Title |
WT_Rpb3_1 (2) |
Sample type |
genomic |
|
|
Channel 1 |
Source name |
IP
|
Organism |
Saccharomyces cerevisiae |
Characteristics |
strain: WT tag: untagged antibodies: Rpb3 (Neoclone: W0012)
|
Treatment protocol |
The cells were grown in SC and induced for Gcn4, by treating cells grown in SC with with 0.65 µM of sulfometuron methyl (SM; Chemservice, cat# N-13254) for 20 minutes and processed for ChIP analysis.
|
Growth protocol |
Chromatin immunoprecipitation experiments were performed as described previously (Govind et al., 2012). Briefly, 100 ml of cells (A600 = 0.6) were cross-linked with 1% formaldehyde for 15 minutes at ambient temperature and quenched with glycine. Chromatin was isolated and fragmented by sonication (Branson450) to an average size of 300-400 base pairs. The soluble fraction of chromatin was used for ChIP using the appropriate antibodies.
|
Extracted molecule |
genomic DNA |
Extraction protocol |
The ChIP and Input DNA was extracted by Phenol:Chloform method after reverse-crosslinking at 65 degree C, and after proteinase K treatment. DNA was fragmented with DNase prior to labeling
|
Label |
Alexa555
|
Label protocol |
BioPrime Array CGH Genomic Labeling Module was used for labeling and the manufacturer's recommended protocol was followed.
|
|
|
Channel 2 |
Source name |
input
|
Organism |
Saccharomyces cerevisiae |
Characteristics |
strain: WT tag: untagged
|
Treatment protocol |
The cells were grown in SC and induced for Gcn4, by treating cells grown in SC with with 0.65 µM of sulfometuron methyl (SM; Chemservice, cat# N-13254) for 20 minutes and processed for ChIP analysis.
|
Growth protocol |
Chromatin immunoprecipitation experiments were performed as described previously (Govind et al., 2012). Briefly, 100 ml of cells (A600 = 0.6) were cross-linked with 1% formaldehyde for 15 minutes at ambient temperature and quenched with glycine. Chromatin was isolated and fragmented by sonication (Branson450) to an average size of 300-400 base pairs. The soluble fraction of chromatin was used for ChIP using the appropriate antibodies.
|
Extracted molecule |
genomic DNA |
Extraction protocol |
The ChIP and Input DNA was extracted by Phenol:Chloform method after reverse-crosslinking at 65 degree C, and after proteinase K treatment. DNA was fragmented with DNase prior to labeling
|
Label |
Alexa647
|
Label protocol |
BioPrime Array CGH Genomic Labeling Module was used for labeling and the manufacturer's recommended protocol was followed.
|
|
|
|
Hybridization protocol |
The samples were lableled using BioPrime Array CGH Genomic Labeling Module kit and hybridized according to the Manufacturer's recommended protocol
|
Scan protocol |
Agilent Technologies Scanner G2505B US45102973
|
Description |
Sulfometuron methyl Biological Replicate-1
|
Data processing |
Data processed on R. Median normalization carried out.
|
|
|
Submission date |
Jan 20, 2016 |
Last update date |
Jul 01, 2016 |
Contact name |
Chhabi Govind |
E-mail(s) |
govind@oakland.edu
|
Organization name |
Oakland University
|
Department |
Biological Sciences
|
Street address |
333 Science and Engineering Building
|
City |
Rochester |
State/province |
mi |
ZIP/Postal code |
48085 |
Country |
USA |
|
|
Platform ID |
GPL10930 |
Series (1) |
GSE77016 |
Recruitment of Saccharomyces cerevisiae Cmr1/Ydl156w to coding regions promotes transcription genome wide |
|