RNA sequencing of the heads of male castes in Nasutitermes takasagoensis by Illumina Hiseq

For RNA-seq analysis, total RNA was extracted from the heads of 50 individuals each of soldier, presoldiers, minor workers and male alates. The heads of soldiers were divided into two parts: a frontal gland Class 1 cell layer and other tissues. As a comparison, we also examined the head tissues (including the frontal gland Class 1 cell layer) in another colony. RNA extraction was performed using an RNeasy mini Kit with DNase treatment. TruSeq RNA sample preparation kits (Version 2) were used to generate a complementary DNA library from 1 ?g of total RNA from each caste or tissue. The cDNA libraries were individually indexed for multiplex sequencing before being pooled and sequenced on a HiSeq 2000 platform with 101 bp paired-end reading.