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RNA-Seq of Ovis aries: adult male fat-tail
After the total RNA extraction and DNase I treatment, magnetic beads with Oligo (dT) were used to isolate mRNA. ixed with the fragmentation buffer, the mRNA fragmented into short fragments. Then cDNA synthesized using the mRNA fragments as templates. Short fragments purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The suitable fragments selected for the PCR amplification as templates. During the QC steps, Agilent 2100 Bioanaylzer and ABI StepOnePlus Real-Time PCR System were used in quantification and qualification of the sample library. At last, the library were sequenced using Illumina HiSeq2000.
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SUB4873173
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