title: Invertebrate sample from Tamalia coweni
design: Tamalia aphid galls were collected from 13 species of host plants at 17 sites across Arizona, Nevada and California (Fig. 1; Table S1; S2). A total of six Tamalia species were collected:  T. coweni (Cockerell), T. dicksoni Remaudière & Stroyan, T. inquilinus Miller, and three novel entities, Tamalia Species A, Tamalia Species B, and Tamalia Species C.  All collections were made between May 2010 and May 2011. Aphids were dissected from fresh galls and preserved immediately at -80°C. Subsequently, aphids within 1.5 mL tubes were immersed in liquid nitrogen and pulverized with a micropestle prior to DNA or RNA extractions. Voucher specimens will be deposited in the Smithsonian Institution Department of Entomology, Canadian National Collection of Insects, Washington State University, and California State University, Chico.      High-throughput sequencing       Total genomic RNA was extracted from four Tamalia samples of 50 adult animals each, representing T. coweni and T. inquilinus populations on the host plants Arctostaphylos viscida and Arctostaphylos manzanita in northern California. Aphid samples were preserved in an ultracold freezer, and separated by age (juveniles and adults) on dry ice.  Adults were pooled from approximately 10 galls, and ground in liquid nitrogen. We used the Qiagen RNeasy Mini Kit® for cells, tissues and yeast, including the DNase digestion protocol. Purified RNA was quantified with a NanoDrop ND-1000 spectrophotometer. Four libraries were constructed and sequenced through a paired-end read protocol on the Illumina Genome Analyzer at the HudsonAlpha Institute for Biotechnology Genome Services Lab (hudsonalpha.org). Illumina instrument software was used to perform data analysis and base calling.