Larval/Parasitic Sea Lamprey Brain
Total RNA was extracted using VersaGene RNA Tissue Kit (Gentra, Inc). The quality of the extracted RNA was verified by gel electrophoresis and quantified using a NanoDrop ND-1000 spectrophotometer. 1ug of total RNA was used as a template for first strand cDNA synthesis using the SMART™ cDNA Synthesis Kit (Clonetech Laboratories, Inc.). 13 cycles of LD-PCR was performed to amplify single-strand cDNA using the Advantage® 2 PCR Kit (Clonetech Laboratories, Inc.) according to manufacturer’s instructions. PCR products were purified using QIAquick PCR Purification Kit (Qiagen, Inc) and concentrated on Millipore YM-30 (MWCO 30,000) columns. 5200ng of cDNA were submitted to MSU RTSF for 454 sequencing.