RNA-Seq of Solanum lycopersicum: Tamaulipas pollen tube at 37C
For each biological replicate pollen from each cultivar was collected in a single 0.5ml conical tube. 300ul of tomato liquid pollen growth medium (tPGM) was added to the tube and mixed to break clumps. 150ul was immediately transferred to a new tube and 300ul tPGM added to both the new and old tubes. Both tubes were then moved to a water bath at 28C. After 3 hours of incubation, 1 tube was moved to 37C (S), the other left at 28C (C), and both incubated for an additional 3 hours. After incubation pollen tubes were collected by centrifugation and stored at -80C. Paired end RNA-Seq libraries were created with the NEBNext Ultra RNA Library Prep Poly A kit per the manufacturers protocol.