title: NextSeq 500 sequencing; Genomic and transcriptomic landscapes of independent invasions of the Pacific Oyster Crassostrea gigas
design: Genomic and transcriptomic landscapes of independent invasions of the Pacific Oyster Crassostrea gigas
library_construction_protocol: The oysters were dissected immediately after collection. The gills were frozen in the liquid nitrogen and then stored at -80°C.  RNA was extracted from 30 mg with TRIzol® Reagent (Life Technologies, Carlsbad, California, US) following the manufacturer's protocol. To homogenize the gill tissue, the reagent was added to the frozen samples together with a 5 mm stainless steal bead and a mixture of 2 mm glass beads. The tubes were then placed in pre-cooled TissueLyzer adapters (Qiagen, Hilden, Germany) and bead-beaten at 30 Hz for 2 x 90 s in a ball mill (Retsch, Haan, Germany). The quality and concentration of the extracted RNA were checked with a NanoDrop ND-1000 spectrophotometer (Peqlab (VWR), Erlangen, Germany) and the RNA was subsequently stored at -80°C.  The dual-indexed libraries were prepared from 1 µg RNA per sample with the TruSeq Stranded mRNA HT Sample Prep Kit (Illumina, San Diego, CA, USA). The quality and concentration of the resulting libraries was checked with an Agilent 2100 Bioanalyzer using the Agilent DNA 7500 Nano Kit (Agilent Technologies, Waldbronn, Germany). All of the above kits were used according to the manufacturers' protocols. Data on DNA fragment length and concentration were then used to calculate the molarity of individual libraries. These were subsequently pooled equimolarly (10 nM)
attributes.tag: Experimental Factor: compound
attributes.value: Mix of ampiciline, gentamicine, tetracycline and kanamycine, each 100 g/l seawater each