|
| Status |
Public on Sep 18, 2012 |
| Title |
wt_pH 7.5-6.5_rep3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
wild type at pH 7.5
|
| Organism |
Synechocystis sp. PCC 6803 |
| Characteristics |
genotype/variation: wild type
strain: ATCC27184
ph: 7.5
|
| Treatment protocol |
For the acid stress experiments, cells of wild type and slr0643 mutant were grown at standard condition before harvesting by centrifugation at 2000g for 5min. The collected cells were then resuspended in fresh BG11 medium containing 20 mM HEPES-NaOH, pH 7.5 or pH6.5.
|
| Growth protocol |
The original Synechocystis sp. PCC 6803 wild type strain was derived from ATCC, namely ATCC27184. Standard growth condition was at 30°C in BG11 medium containing 20 mM HEPES-NaOH (pH 7.5) under continuous illumination of 30 μmol m−2 s−1, with oscillation at 130rpm. Kanamycin at 40ug mL-1 was supplemented in BG11 medium for slr0643 mutant.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were broken in Mini-Bead Beater-1 (Biospec, USA) and total RNA was extracted using RNA extraction kit (DongSheng Biotech, China). RNase-free DnaseⅠ (TAKARA Biotech, China) was used to remove contaminant of genomic DNA.
|
| Label |
Cy5
|
| Label protocol |
Dye-labeled cDNA was synthesized using RNA Fluorescence Labeling Core Kit (M-MLV Version) Ver.2.0 (TAKARA Biotech, China).
|
| |
|
| Channel 2 |
| Source name |
wild type at pH 6.5
|
| Organism |
Synechocystis sp. PCC 6803 |
| Characteristics |
genotype/variation: wild type
strain: ATCC27184
ph: 6.5
|
| Treatment protocol |
For the acid stress experiments, cells of wild type and slr0643 mutant were grown at standard condition before harvesting by centrifugation at 2000g for 5min. The collected cells were then resuspended in fresh BG11 medium containing 20 mM HEPES-NaOH, pH 7.5 or pH6.5.
|
| Growth protocol |
The original Synechocystis sp. PCC 6803 wild type strain was derived from ATCC, namely ATCC27184. Standard growth condition was at 30°C in BG11 medium containing 20 mM HEPES-NaOH (pH 7.5) under continuous illumination of 30 μmol m−2 s−1, with oscillation at 130rpm. Kanamycin at 40ug mL-1 was supplemented in BG11 medium for slr0643 mutant.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were broken in Mini-Bead Beater-1 (Biospec, USA) and total RNA was extracted using RNA extraction kit (DongSheng Biotech, China). RNase-free DnaseⅠ (TAKARA Biotech, China) was used to remove contaminant of genomic DNA.
|
| Label |
Cy3
|
| Label protocol |
Dye-labeled cDNA was synthesized using RNA Fluorescence Labeling Core Kit (M-MLV Version) Ver.2.0 (TAKARA Biotech, China).
|
| |
|
| |
| Hybridization protocol |
Hybridization was conducted at 65 1C for 16 h. After incubation, the microarrays were rinsed with 2 x SSC (1 x SSC is 150 mM NaCl and 15 mM sodium citrate) at room temperature, with 2 x SSC at 60 1C for 10 min, with 0.2 x SSC, 0.1% SDS at 60 1C for 10 min, and finally with distilled water at room temperature for 2 min. Moisture was removed with an air spray prior to analysis in the array scanner
|
| Scan protocol |
Images were acquired with GMS418 array scanner (Affimetrix, USA)
|
| Data processing |
Scanned images were quantified by ImaGene V.4.2 software (BioDiscovery, USA). Spot signal intensity was determined after local background subtraction and normalization
|
| |
|
| Submission date |
May 03, 2012 |
| Last update date |
Sep 18, 2012 |
| Contact name |
Gu CHEN |
| E-mail(s) |
[email protected]
|
| Organization name |
South China University of Technology
|
| Street address |
381 Wushan Road
|
| City |
Guangzhou |
| ZIP/Postal code |
510641 |
| Country |
China |
| |
|
| Platform ID |
GPL15530 |
| Series (1) |
| GSE37747 |
Slr0643, a S2P Homolog, Is Essential for Acid Acclimation in Cyanobacterium Synechocystis sp. PCC 6803 |
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