Small-RNA libraries were generated from the nine samples of Yellow River carp: primordial gonad control (PG-CK), primordial gonad exposed to atrazine for 8 h (PG-A8h), primordial gonad exposed to atrazine for 24 h (PG-A24h), juvenile ovary control (IIC-CK), juvenile ovary exposed to atrazine for 8 h (IIC-A8h), juvenile ovary exposed to atrazine for 24 h (IIC-A24h), juvenile testis control (IIX-CK), juvenile testis exposed to atrazine for 8 h (IIX-A8h), juvenile testis exposed to atrazine for 24 h (IIX-A24h). Small-RNA libraries were generated using the mirVanaTM mircoRNA Isolation Kit (Ambion, USA), according to the manufacturer’s instructions. Small-RNA libraries were prepared from three biological replicates for each sample.
Total RNA was ligated with 3′ and 5′ RNA adaptors. Fragments with adaptors on both ends were enriched by PCR after reverse transcription, as described previously [49 (link)]. The resulting cDNAs were purified and enriched with 6% denaturing polyacrylamide gel electrophoresis to isolate the fractions of the expected size and to eliminate unincorporated primers, primer dimer products, and dimerized adaptors [49 (link)]. Finally, the nine resulting RNA libraries were sequenced using an Illumina/Solexa Genome Analyzer, at Guangzhou Genedenovo Biotech Company (Guangzhou, China).
Total RNA was ligated with 3′ and 5′ RNA adaptors. Fragments with adaptors on both ends were enriched by PCR after reverse transcription, as described previously [49 (link)]. The resulting cDNAs were purified and enriched with 6% denaturing polyacrylamide gel electrophoresis to isolate the fractions of the expected size and to eliminate unincorporated primers, primer dimer products, and dimerized adaptors [49 (link)]. Finally, the nine resulting RNA libraries were sequenced using an Illumina/Solexa Genome Analyzer, at Guangzhou Genedenovo Biotech Company (Guangzhou, China).
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