title: RNAseq of Glycine max: infected with Rhizoctonia solani AG1 IA sample 1
design: Total RNA (10 ľg) from infected and control samples at the 24 h time point was used to isolate mRNA for RNA sequencing (RNAseq).   RNAseq library preparation was performed following modified methods of Kumar et al. (2012).  Briefly, mRNA was isolated using DynaBeads¨ (ThermoFisher Scientific), and first and second strand cDNA synthesis was done following the Kumar et al. (2012) protocols with AMPure¨ bead purification (Beckman Coulter).  cDNA was fragmented using Fragmentase (NEB) with incubation at 37űC for 20 minutes.  End repair and A-tailing were performed using the NEB enzyme mix and Klenow 3Ő to 5Ő exo (ThermoFisher Scientific).  Adapter ligation was done using 2X rapid T4 DNA ligase (NEB) and NEXTflex-96 RNA-seq barcode adapters 85 to 96 (BiooScientific,).  Samples were then PCR enriched using 1X Phusion High Fidelity PCR Master Mix (NEB), 0.67 ľM each primer PE1/PE2 (BiooScientific) and 7.5 ľL of the sample with the following thermocycle conditions found in Kumar et al. (2012) for a total of 15 cycles
library_name: Soybean_RFB_4