|
| Status |
Public on Mar 14, 2013 |
| Title |
RNA_WT_36C |
| Sample type |
RNA |
| |
|
| Source name |
Total RNA, WT, 36C
|
| Organism |
Schizosaccharomyces pombe |
| Characteristics |
strain: Hu303
genotype/variation: WT
temperature: 36C
|
| Growth protocol |
Liquid cultures were grown in YES media shaking at 180 rpm. The temperature sensitive top3-105 mutant and wild type controls were grown at the permissive temperature (25 °C) over night and then for 8 hours at the nonpermissive temperature (36°C) before harvesting. All other strains were grown at 30 °C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from mid-logarithmic phase cells after 8 hours at 36C using the hot-phenol method. Briefly, cells were harvested by centrifugation at RT, resuspended in TES (10 mM TrisHCl pH7.5, 10 mM EDTA and 0.5% SDS) and transferred to preheated acid phenol (65 °C). After 60 min shaking incubation at 65 °C, RNA was isolated by phenol-chloroform extraction.
|
| Label |
biotin
|
| Label protocol |
RNA was treated according to the Affymetrix total RNA labelling protocol (http://www.affymetrix.com).
|
| |
|
| Hybridization protocol |
Hybridised to Affymetrix GeneChip S. pombe Tiling 1.0FR Affymetrix core facility at Novum (http://apt.bea.ki.se) according to standard Affymetrix protocols.
|
| Scan protocol |
Scanned at the Affymetrix core facility at Novum (http://apt.bea.ki.se) according to standard Affymetrix protocols.
|
| Description |
Total RNA from WT cells after 8 hours at 36C
RNA_WT_36C
|
| Data processing |
CEL.files were analysed using Affymetrix Tiling Analysis Software (TAS) v1.1. One sample analysis of replicates A+B and linear scaling to 100 was applied. Linear probe signals were generated using a bandwidth of 100 and assigned to S. pombe genome coordinates (Sanger 2007) and reported in BAR files (Fwd and Rev strands).
|
| |
|
| Submission date |
Feb 08, 2013 |
| Last update date |
Mar 14, 2013 |
| Contact name |
Karl Ekwall |
| E-mail(s) |
karl.ekwall@ki.se
|
| Phone |
+46 8 6089133
|
| Organization name |
Karolinska Inst
|
| Street address |
Alfred Nobels Alle 7
|
| City |
Stockholm |
| ZIP/Postal code |
S-141 89 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL7715 |
| Series (1) |
| GSE44206 |
DNA topoisomerase III localizes to centromeres and affects centromeric CENP-A levels in fission yeast |
|
