Seeds were surface sterilized and grown on MS plates for 10-14 days before transfer to a mixture of soil (ED73, Universalerde, Germany) and sand (5:1) fertilized with Plantomaag (Syngenta, Switzerland) and Osmocote (Scotts, Marysville, USA). Plants were grown in a greenhouse chamber with 60% humidity and 16 h light/ 8 h darkness at 20 °C and 16 °C, respectively.
Extracted molecule
total RNA
Extraction protocol
For total RNA isolation from cells or tissues isolated by laser assisted microdissection, caps were covered with 10 ul extraction buffer from the PicoPure RNA isolation kit (Arcturus Engineering, Mountain View, USA), incubated at 42°C for 30 min before isolation according to manufacturers instructions. The DNAse treatment was performed on-column using the RNAse-free DNase set (QIAGEN, Hilden, Germany).
Label
biotin
Label protocol
During the second round of amplification with the Message AmpII Kit (Ambion), biotin-11-UTP (ambion) was incorporated in th amplified aRNA for microarray analysis.
Hybridization protocol
Hybridization-EukGE-WS2v4
Scan protocol
GCOS
Description
RNA amplification with Ambion Message AmpII Kit synergid_cells1
Data processing
BgPANP algorith with R/Bioconductor presence absence p-values with BgPANP
