|
| Status |
Public on Feb 03, 2021 |
| Title |
Rrm3-Flag |
| Sample type |
genomic |
| |
|
| Source name |
G2/M arrest with Nocodazole
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: HY4916
strain genotype: W303 MATa Rrm3-10Flag::KANMX4
antibody: Anti-FLAG M2 F1804 (SIGMA)
|
| Treatment protocol |
G2/M synchronization was performed by 10-20 μg/ml Nocodazole for 120-180 minutes at 25°C and validated by microscopy. Cells were treated with 0.2 mg/ml of Auxin (stock 50 mg/ml) during the arrest, if mentioned. Cells were collected and fixed with 1% formaldehyde for 30 min at room temperature for further ChIP.
|
| Growth protocol |
Cells were grown in YP+Glucose 2% (YPD) media at 25°C in 500 ml flask for a density of 1.2×10^7 cells/ml.
|
| Extracted molecule |
genomic DNA |
| Extraction protocol |
Binding profiles by ChIP-on-chip were determined as described in (Bermejo et al., 2009).
|
| Label |
Dynabeads Protein A invitrogen
|
| Label protocol |
Sample preparation and labeling were carried out following the Affymetrix GeneChip Expression Anaysis protocol.
|
| |
|
| Hybridization protocol |
Labelled probes were hybridized to Affymetrix S.cerevisiae whole genome tiling R arrays.
|
| Scan protocol |
Arrays were scanned on an Affymetrix Scanner.
|
| Description |
Binding pattern of Rrm3 by ChIP-on-chip after arrest of cells in G2/M with Nocodazole
|
| Data processing |
BAR files were produced with Tiling Array Suite 1.1 from the command line with these options: tilecore -analysis -bpmap Sc03b_MR_v04.bpmap -type 1 -n 1 -nt 1 -export 1 -output_type 0 -format_type 1 -band 150 -pval_scale 0 -sig_scale 2 -tail_type 1 -nscale 1 -tgt 500 [files]
|
| |
|
| Submission date |
Mar 24, 2020 |
| Last update date |
Feb 03, 2021 |
| Contact name |
Dana Branzei |
| E-mail(s) |
dana.branzei@ifom.eu
|
| Organization name |
IFOM Foundation - The FIRC Institute of Molecular Oncology Foundation
|
| Lab |
DNA Repair lab
|
| Street address |
Via Adamello 16
|
| City |
Milan |
| ZIP/Postal code |
20139 |
| Country |
Italy |
| |
|
| Platform ID |
GPL7250 |
| Series (1) |
| GSE147452 |
Smc5/6 functions with Sgs1-Top3-Rmi1 to complete chromosome replication at natural pause sites |
|
