RNA-seq of Apis mellifera: hygienic behavior
Libraries were prepared with TruSeq RNA sample preparation kit (Illumina) at the Iowa State University DNA Facility. Total RNA was purified to obtain the mRNA, using polyT oligoattached magnetic beads, and then fragmented it. RNA fragments primed with random hexamers was reverse transcribed into first strand cDNA using reverse transcriptase (SuperScript II) and random primers. Then the cDNA was converted into double stranded DNA and subjected to library preparation. The 10-dsDNA samples were attached to adapters for hybridization onto a flow cell and amplified by PCR. The quality of the libraries was controlled by an Agilent Technologies 2100 Bioanalyzer with Agilent DNA1000 kit. Before sequencing, all samples were normalized and pooled together using the adapter indices provided in the kit.  Samples were sequenced in single-end read mode (1x), with 100 bp reads, on a HiSeq 3000 machine (Illumina).