title: India Andaman Island animals
organism: Penaeus monodon
description: Three individuals from Andamans: 11° 40' N &amp; 92° 45' Shrimp were dissected under sterile conditions and fresh tissue samples (hepatopancreas, muscle, nervous tissue, intestinal tract, heart, gonad, gill, pleopod and lymphoid organ) were collected and stored in 1.5 ml microtubes. Microtubes were filled with RNAlater (Qiagen) to immerse the entire tissue. Subsequently, tubes were topped up to the brim with RNAlater, capped and stored at room temperature for two hours, 4 oC overnight and -80 oC until further use. RNA was extracted (TRIzol® Plus RNA Purification, Invitrogen) according to manufacturer’s instructions. Tissue was ground in liquid nitrogen and 1.5 ml of TRIzol was added. RNA was further purified by chloroform extraction followed by precipitation using isopropanol and one wash with 70% ethanol. RNA was resuspended in 25 µl of RNase-free water. Only samples with optimal purity (OD260/280 &gt;1.8 and &lt;2.2; OD260/230&gt;1.0 and &lt;2.4) and concentration (&gt;50 ng/µl and &lt;2500 ng/µl), estimated using UV spectrophotometry (Nanodrop) and capillary electrophoresis (Agilent Bioanalyser), were utilized for sequencing. Samples were prepared using the Illumina protocol outlined in “Preparing Samples for Sequencing of mRNA” (Part# 1004898 Rev. A September 2008).