7 protocols
AccessionType
image_acquisition
Agilent G2565AA and G2565BA Microarray Scanner and Feature ExtractionVersion: 7.0,10.5Agilent publication number: G2566-90017, G4460-90019URL: http://www.chem.agilent.com/Library/usermanuals/Public/G2566-90017.pdf, http://www.chem.agilent.com/Library/usermanuals/Public/G4460-90019_FE_10.5_User.pdfScanner manual for software release 7.0. The Feature Extraction Software User Guide shows you how to set up and run Feature Extraction automatically for a batch of image files and how to extract image files in real time.
nucleic acid hybridization to array protocol
Overall 25,262 transcripts were represented on the microarray by 1, 2 or 3 individual probes. Hybridizations for all experimental conditions were performed in 4 replicates. Total RNA samples derived from the treatments were hybridized against the pooled control respective to their origin. The microarray hybridization procedure was carried out with 300ng of cyanine-3 and cyanine-5 labelled cRNA for 17h at 65°C. Control/control hybridization were performed, each component of the pooled control (LP 2°C, LP 7°C, LP 12°C) was hybridized against the pooled control to mitigate dye bias effects. Subsequently microarray disassembly and wash procedure followed as described by the manufacturer’s instructions (Agilent).
labeling
The labeling experiment was performed according to the manufacture's protocol (Agilent technologies, Low RNA Input Fluorescent Linear Amplification Kit Protocol, Version 5.0.1).
(Parameters: Amplification = none, Mass unit = Micro gram)
labeling
The labeling experiment was performed according to the manufacture's protocol (Agilent technologies, Low RNA Input Fluorescent Linear Amplification Kit Protocol, Version 5.0.1).
(Parameters: Amplification = none, Mass unit = Micro gram)
grow
The growth conditions of the plants are as follows: The plants were grown in the plastic dishes (20 plants/plastic dish) containing germination medium (GM). GM contained Murashige and Skoog salts, 1% sucrose (WAKO, Osaka, Japan) and 0.9% Bacto-agar (Difco, Detroit, MI, USA). The plants were grown for 3 weeks in a growth chamber at 22C and 50-60% humidity under 16 hr light/8 hr dark and then subjected to the stress treatments.
(Parameters: time unit = seconds, temperature unit = C)
nucleic_acid_extraction
Total RNAs were isolated using the RNAiso Reagent (Takara, Japan).
(Parameters: Extracted product = total_RNA, Amplification = none)
specified_biomaterial_action
The plants were incubated at 22C under non-stress condition.