RNA-Seq analysis of synchronised developing pollen isolated from a single anther

To assist with understanding gene expression during pollen development we have developed a protocol to perform RNA-Seq on pollen isolated from a single anther (SA RNA-Seq). The protocol involves removing pollen from a single anther for analysis and viewing the remaining pollen to determine the developmental stage. The isolated pollen is chemically lysed, mRNA isolated from the lysate using an oligo-dT column before library preparation. We have demonstrated the efficacy of this approach in the model species Arabidopsis thaliana and the fruit crop kiwifruit (Actinidia chinensis)