Illumina HiSeq 2000 sequencing; Elodea nuttallii transcriptomic response to mercury, monomethyl-mercury, copper and UV-B radiation
Elodea nuttallii transcriptomic response to mercury, monomethyl-mercury, copper and UV-B radiation
Extract 16
Elodea nuttallii were grown in 25-L aquariums supplemented with a layer of sediment in a 16/8 h light/dark cycle at 20  degrees C. 3 shoots per treatment were cut and exposed in 1-L PE beakers to 0 (control), 20, 200 2000, 20000 or 200000 ng/L HgCl2, 10, 100, 1000 or 10000 ng/L methyl-Hg, or 10-6 M CuCl2 for 24 h, or to 51 uW/cm2 UV-B radiation for 6 h. Double exposures were conducted for UV-B with 200 or 200000 ng/L HgCl2. All treatments except 20 and 20000 ng/L HgCL2, 10, 100 and 1000 ng/L methyl-Hg and 10-6 M Cu were run in biological triplicate. Shoots were ground in liquid nitrogen and RNA was extracted using Trizol reagent. Libraries were constructed at Fasteris SA (Geneva, CH) using the Illumina TruSeq stranded mRNA kit according to the manufacturer's instructions. Briefly, mRNA was purified and fragmented, and subsequently first and second cDNA strands were synthesised. Finally, after adapter ligation, libraries were PCR amplified and quality checked.