Regulatory control of high levels of carotenoid accumulation in potato tubers
Hybridization: POR03/Atlantic |
| Hybridization protocol |
| Hybridization solution | Genisphere Hybridization Solution |
| Solution concentration | 0.25m NaPO4, 4.5%SDS, 1mm EDTA, 1x SSC, 2x Denhardt's Solution |
| Blocking agent | ssDNA and Oligo dT |
| Wash procedure | 3x15 minutes: 1) 2x SSC + 0.2% SDS at 60 C. 2) 2x SSC at 22 C. 3) 0.2x SSC at 22 C |
| Labeled Target Quantity | 3 ul cDNA |
| Hybridization Time | 12-16 hours |
| Hybridization Volume | 60 ul |
| Hybridization Temperature | 60C |
| Hybridization Instrument | 50 ml tube in 60C chamber |
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| Scan information |
| Scan hardware | Scan Array 5000 |
| scan software | Scan Array ver 3.1 |
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| Image analysis information |
| Software | Affymetrix GeneChip Operating Software (GCOS) |
| Analysis algorithm | MAS 5 software |
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| Normalization information |
| Normalization software | marray package under R |
| Normalization algorithm | Print-tip specific Lowess Normalization |
| Normalization strategy | Total Array |
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