cell type: Tapetal Cell developmental stage: 1.8-2.0 mm anther
Growth protocol
Maize ( Zea mays ) B73 inbred line plants were grown in the greenhouse under 14h light and 10h dark in a growth chamber at 22-26℃ with 65% relative humidity.
Extracted molecule
total RNA
Extraction protocol
Total RNA from Laser microdissection samples were extracted with the PicoPure RNA isolation kit (Applied Biosystems), Amino-ally1 aRNA was recovered through 2-round amplification using a Target Amp™ 2-Round Aminoallyl-aRNA Amplification Kit.
Label
cy5
Label protocol
Total RNA were labeled by Cy3 NHS ester (Cat#PA13105, GE healthcare Biosciences, Pittsburgh, PA, US) ，Cy5 NHS ester (Cat#PA15100, GE healthcare Biosciences, Pittsburgh, PA, US)followed the manufacturer’s instructions. Labeled cRNA were purified by RNeasy mini kit (Cat#74106, QIAGEN, GmBH, Germany).
Hybridization protocol
Each Slide was hybridized with 60pmol Cy3-labeled cRNA and 30pmol Cy-5 labeled cRNA in Hybridization Oven. After 17 hours hybridization, slides were washed in staining dishes.
Scan protocol
Slides were scanned by Agilent Microarray Scanner (Cat#G2565BA, Agilent technologies, Santa Clara, CA, US)and Feature Extraction software 10.7 (Agilent technologies, Santa Clara, CA, US) with default settings. Raw data were normalized by Lowess(locally weighted scatter plot smoothing) algorithm, Gene Spring Software 11.0 (Agilent technologies, Santa Clara, CA, US).
