RADseq of kalotermes approximatus worker
Samples were stored in 96% ethanol at 4C until DNA extraction. Total genomic DNA was extracted from each individual using a modified Gentra Puregene extraction method (Gentra Systems, Inc. Minneapolis, MN, USA). DNA quality and concentration were assessed by agarose gel electrophoresis and Qubit 2.0 Fluorometer (Invitrogen, USA). Suitable genomic DNA was used to construct ddRAD libraries. Libraries were prepared and sequenced at the Texas A&M AgriLife Genomics and Bioinformatics Service facility using SphI and EcoRI restriction enzymes following the protocol of Peterson et al. (2012). Each sample was identified using a unique indexed barcode. Samples were amplified through PCR with iProof High-Fidelity DNA Polymerase (Bio-Rad). PCR products were purified using AMPure XP beads (Beckman Coulter Inc.). Libraries were size-selected to a range of 300500 bp using the BluePippin system (Sage Science Inc.). Libraries were sequenced on six flowcell lanes using an Illumina HiSeq 2500 (Illumina Inc., USA) to generate 150 bp pairend reads.