title: Skin transcriptome
design: Six sibling Oujiang color common carp samples were collected, including three Fenyu (completely white) and three Dahua (red with big black spots). Total RNA of skin tissues was isolated using TRIzol reagent (Invitrogen) following the standard protocol. Total RNA of fish with the same color pattern was pooled and incubated with 10 U DNase I (Ambion) at 37ºC for 1 hr, and then nuclease-free water was added to bring the sample volume to 250 μL. Double-stranded cDNA was synthesized from mRNA according to Ng's full-length cDNA synthesis protocol with minor modifications (Ng et al., 2005). A GsuI-oligo dT primer was used to prime first-strand cDNA synthesis from 10μg of mRNA, using 1000 units of Superscript II reverse transcriptase (Invitrogen). After incubation at 42ºC for 1 hr, the 5’-CAP structure of mRNA was oxidized by NaIO4 (Sigma) and ligated to biotin hydrazide, which was used to select complete mRNA/cDNA by binding Dynal M280 beads (Invitrogen). After the second strand cDNA synthesis, the polyA and 5’ adaptor were removed through GsuI digestion.
library_name: Oujiang common carp