|
| Status |
Public on Nov 05, 2004 |
| Title |
bim1D dSLAM UpTag experiment1 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Pool of haploid single YKOs result from ura3D::URA3 transfromation and sporulation
|
| Organism |
Saccharomyces cerevisiae |
| Extracted molecule |
total RNA |
| |
|
| Channel 2 |
| Source name |
Pool of haploid double YKOs result from bim1D::URA3 transformation and sporulation
|
| Organism |
Saccharomyces cerevisiae |
| Extracted molecule |
total RNA |
| |
|
| |
| Description |
A pool of 5,896 haploid-convertible heterozygous diploid yeast knockout (YKO) strains were transformed with a ura3D::URA3 (as target source1) or a bim1D::URA3 (as target source2) cassette.
The resultant pools of Ura+ transformants were sporulated and spores were spread on MM-Ura.
Pools of Ura+ haploid YKOs were harvested after incubation at 30 degree for 2 days.
Genomic DNA samples were prepared from these two haploid pools and used as the templates for PCR amplification of the TAGs identifying the YKOs with a pair of primers universal for either Uptags or Downtags.
The PCR products from sample1 were labeled with Cy5 and those from sample2 with Cy3.
The Cy5- and Cy3-labeled Uptags and Downtags were mixed together and hybridized to a Rosetta_UpTag.
Fluorescent array images were collected for both Cy5 and Cy3 with a GenePix4000B microarray scanner.
The image data was annalyzed with the Imagene 5.0 and Genesight 3.1 programs from Biodiscovery.
Keywords = bim1D, dSLAM, heterozygotes, YKO
|
| |
|
| Submission date |
Sep 13, 2004 |
| Last update date |
May 27, 2005 |
| Contact name |
Xuewen Pan |
| E-mail(s) |
xpan2@jhmi.edu
|
| Phone |
410-955-2477
|
| Organization name |
Johns Hopkins University School of Medicine
|
| Department |
Department of Molecular Biology and Genetics
|
| Lab |
Boeke Lab
|
| Street address |
733 N. Broadway, Room 331
|
| City |
Baltimore |
| State/province |
MD |
| ZIP/Postal code |
21205 |
| Country |
USA |
| |
|
| Platform ID |
GPL1446 |
| Series (2) |
|
| Data table header descriptions |
| ID_REF |
Serial identifier for probe |
| F635_Median |
Cy5 (F635) median fluorescence intensity |
| B635_Median |
Cy5 (F635) background median fluorescence intensity |
| F532_Median |
Cy3 (F532) median fluorescence intensity |
| B532_Median |
Cy3 (F532) background median fluorescence intensity |
| F635-B635_Median |
Background substracted Cy5 (F635) median fluorescence intensity |
| F532-B532_Median |
Background substracted Cy3 (F532) median fluorescence intensity |
| VALUE |
log ratio (log2 of PRE_VALUE) |
| Flags |
|
| PRE_VALUE |
Normalized ratio defined by F635-B635_Median/F532-B532_Median |
