genotype: Grass phenotype: Water stressed accession: 090729_Sstaph_MO_EXP tissue: leaf
Growth protocol
Sporobolus stapfianus plants were grown on 1 gallon pots filled with ProMix potting soil and alowed to grow for three months under green house conditions. Watering and fertilization were provided as needed and water content measurements were conducted at the time of havest. Leaves were harvested at fully hydrated state, four dehydrating states, a desiccated state, a 12 h rehydrating state, and a 24 h rehydrating state. Leaves were then grinded and used for RNA extraction.
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted using the RNeasy Mini kit (Qiagen Inc., Valencia, CA, USA) and DNA was removed by on-column DNase digestion with the RNase-Free DNase set (Qiagen). After gel elecrophoresis analysis on formaldehyde-containing 1.2% agarose gel, RNA was quantified using an NanoDrop (NanoDrop Technologies, Inc., Rockland, DE).
Label
Cy3
Label protocol
Labeling was performed by NimbleGen Systems Inc., Madison, WI USA, following their standard operating protocol. See www.nimblegen.com.
Hybridization protocol
Hybridization was performed by NimbleGen Systems Inc., Madison, WI, USA following their standard operating protocol. See www.nimblegen.com.
Scan protocol
Scanning was performed by NimbleGen Systems Inc., Madison, WI USA, following their standard operating protocol. See www.nimblegen.com.
Description
2C
Data processing
The raw data (.pair file) was subjected to RMA (Robust Multi-Array Analysis; Irizarry et al. Biostatistics 4(2):249), quantile normalization (Bolstad et al. Bioinformatics 19(2):185), and background correction as implemented in the NimbleScan software package, version 2.4.27 (Roche NimbleGen, Inc.).
