|
| Status |
Public on May 10, 2010 |
| Title |
K42L_replicate2 |
| Sample type |
RNA |
| |
|
| Source name |
Saccharomyces cerevisiae expressing mutant histone H3 protein
|
| Organism |
Saccharomyces cerevisiae |
| Characteristics |
strain: JPY12
genome/variation: expressing plasmid borne histone H3 protein encoding the substitutions K42L
|
| Biomaterial provider |
Boeke Lab, Johns Hopkins University School of Medicine
|
| Treatment protocol |
Site directed mutagenesis was employed to change amino acid number 42 from lysine to leucine on an plasmid borne (pEMH7) histone H3 protein.
|
| Growth protocol |
Cells were grown in rich YPD media to log phase and then harvested.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Hot phenol followed by RNeasy Kit (QIAGEN)
|
| Label |
Biotin
|
| Label protocol |
One round amplification protocol for total RNA following Affymetrix’ specifications using T7 promoter oligo dT primer for cDNA synthesis, and ENZO IVT kit (www.affymetrix.com, GeneChip Expression Analysis, Technical Manual, 2003).
|
| |
|
| Hybridization protocol |
16hrs at 45° C with rotation (60rpm) as described by Affymetrix in their GeneChip Expression Analysis Technical Manual, 2004.
|
| Scan protocol |
Using Affymetrix’ GeneChip Scanner 3000 7G and default parameters described by the manufacturer in their GeneChip Expression Analysis Technical Manual, 2004.
|
| Description |
Hybridization was performed by the Johns Hopkins Medical Institutions Microarray Core Facility.
|
| Data processing |
Image analysis using GCOS 1.4, using MAS 5.0 algorithm and manufacturer’s specifications. Global scaling of images to a target intensity of 150.
|
| |
|
| Submission date |
Dec 09, 2008 |
| Last update date |
May 10, 2010 |
| Contact name |
Edel Maria Hyland |
| Fax |
4105021872
|
| Organization name |
Johns Hopkins School of MEdicine
|
| Department |
Molecular Biology and Genetics
|
| Lab |
Jef Boeke
|
| Street address |
733 North broadway, Rm 320
|
| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21205 |
| Country |
USA |
| |
|
| Platform ID |
GPL2529 |
| Series (1) |
| GSE13889 |
Gate-Keeping transcription through histone H3 K42, a novel site of methylation in the yeast nucleosome core |
|
