RNAseq of stamens  and carpels from Mimulus guttatus IM62, M. nasutus SF5, and sterile (STE) and  fertile (FER) SF5-IM62 introgression hybrids
For this study, we generated 24 RNAseq  libraries representing three biological replicates each of two tissue types  (stamens and carpels) from four genotypes that vary at hms1 and hms2 (M.  guttatus IM62, M. nasutus SF5, and fertile (FER) and sterile (STE) SF5-IM62  introgression progeny. To collect sufficient tissue for each biological  replicate, we dissected eight to 24 pre-anthesis floral buds and transferred  carpel and stamen tissue into separate 1.5 mL microcentrifuge tubes partially  submerged in liquid nitrogen. We extracted RNA using a QuickRNA Miniprep Kit  (Zymo Research, Irvine, CA, USA) then measured RNA concentration using a  Qubit RNA BR (Broad-Range) Assay Kit and a Qubit 2.0 Fluorometer (Thermo  Fisher Scientific Inc., Waltham, MA, USA). We shipped RNA samples overnight  on dry ice to the Duke Center for Genomic and Computational Biology (Durham,  NC, USA), where the Sequencing and Genomic Technologies core checked RNA  quality using an Bioanalyzer 2100 (Agilent Technologies, Santa Clara, CA, USA),  constructed sequencing libraries using a KAPA Stranded mRNA-Seq Kit (F.  Hoffmann-L Roche, Basel, Switzerland), and sequenced all 24 libraries on a  single lane of HiSeq 4000 (Illumina, Inc. San Diego, CA, USA), producing 50bp  single-end reads.