After extracting the total RNA from HCC and cirrhosis tissues, the non-coding RNAs were enriched by removing rRNA from the total RNA with kit. By using the fragmentation buffer, the non-coding RNAs were fragmented into short fragments (about 200~500nt), then the first-strand cDNA was synthesized by random hexamer-primer using the fragments as templates, and dTTP was substituted by dUTP during the synthesis of the second strand. Short fragments were purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments were connected with adapters, then the second strand was degraded using UNG (Uracil-N-Glycosylase) finally. After agarose gel electrophoresis, the suitable fragments were selected for the PCR amplification as templates. During the QC steps, Agilent 2100 Bioanalyzer and ABI StepOnePlus Real-Time PCR System were used in quantification and qualification of the sample library. As last, the library based on mRNA and lncRNA could be sequenced using Illunima HiSeqTM 2000.

Illumina HiSeq 2000 paired end sequencing; HCC_BJ21_non_Cancer_RNA

BJ21B

To conduct clinical cancer research and study the genetic basis of hepatocellular carcinoma

Transcriptome sequencing identified hub genes for hepatocellular carcinoma by weighted-gene co-expression analysis

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Samples