title: Illumina Genome Analyzer IIx sequencing; RNA-seq of N. gansuense-infected (E+) vs. non-infected (E-) A. inebrians
design: RNA-seq of N. gansuense-infected (E+) vs. non-infected (E-) A. inebrians
library_construction_protocol: Both E+ and E- A. inebrians seeds were incubated at 5, 10, 15, 20 and 25 oC in Petri dishes (15-cm diameter). After imbibition, germination rate of seeds was monitored every 24 h during 24-192 h. The seeds showing radical extension of 1 mm were scored and removed from the Petri dishes. After 25 d, when no additional seeds had germinated for 5 d, the germination experiment was terminated. Each treatment was repeated five times independently with 100 seeds in each replicate. Both E+ and E- A. inebrians seeds were sown on filter paper and grown at 10 oC in constant temperature incubators for two days. Total RNAs were isolated from about 10 g of either E+ or E- drunken horse grass seeds using the TRIzol reagent (Invitrogen) and incubated with DNase I (Takara) to remove residual genomic DNA according to manufacturers instruction. The concentration and quantification of all RNA were determined spectrophotometrically.
attributes.tag: Experimental Factor: Non-infected
attributes.value: infect