NextSeq 500 sequencing; Transcription profiling of Arabidopsis thaliana roots and leaves in reaction to fungal (Laccaria bicolor) volatiles
Transcription profiling of Arabidopsis thaliana roots and leaves in reaction to fungal (Laccaria bicolor) volatiles
Leaves/roots were cut off and directly frozen in liquid nitrogen. Arabidopsis thaliana plants were grown from seeds on ½ MS medium in squared Petri dishes, which were placed vertically in a growth chamber. Plants were kept under long-day conditions (16 h light, 120-130 µE/ 8 h dark) at 24 °C during light and 22 °C during darkness. Humidity was kept between 60-80%. L. bicolor was grown from culture on cellophane and P20 medium for 14 days at 22 °C in darkness. Per bi-compartmented squared Petri dish, 15 4-days-old A. thaliana seedlings were co-incubated with 4 14-days-old L. bicolor plugs. Control seedlings were grown with cellophane and P20. Arabidopsis RNA was extracted using RNAeasy extraction kit (Qiagen) according to the manufacturer's protocol. Poly-A RNA was enriched using the NEBNext Poly (A) mRNA Magnetic Isolation Module (New England Biolabs (NEB), Ipswich, Massachusetts, USA). Sequencing libraries were prepared using the NEBNext Ultra RNA Library Prep Kit for Illumina (NEB) according to manufacturer's instructions.