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用氯化铯-异硫氰酸胍超速离心法（CsCl2-GTC）提取肝检组织总RNA，用mRNA提取试剂盒分离mRNA。将分离得到的mRNA在逆转录酶存在的条件下，合成cDNA的第一链；在大肠杆菌DNA聚合酶Ⅰ和RNaseH存在的条件下，合成cDNA的第二链。在引物设计上，引进M13M4引物及EcoRⅠ酶切位点，同时含有Olygod（T）8。为了验证合成的cDNA文库的可靠性，我们设计了几对引物用于PCR扩增TPO和IGF－1基因。实验结果表明，PCR有效地扩增分离出TPO和IGF－1基因。经酶切鉴定和DNA序列分析证实，该序列与文献报道完全一致。该cDNA文库的构建成功，为今后分离和克隆肝细胞产生的细胞因子打下了基础；同时也为从分子水平上研究肝细胞的生物学功能提供了重要材料。
Total RNA was extracted from hepatic tissue by CsCl-guanidine-isothiocyanate ultracentrifugation (CsCl2-GTC), mRNA was isolated by mRNA extraction kit. The isolated mRNA is synthesized in the first strand of cDNA in the presence of reverse transcriptase, and the second strand of cDNA is synthesized in the presence of E. coli DNA polymerase I and RNaseH. Primer design, the introduction of M13M4 primers and EcoR Ⅰ restriction sites, containing Olygod (T) 8. In order to verify the reliability of the synthesized cDNA library, we designed several pairs of primers for PCR amplification of the TPO and IGF-1 genes. The experimental results show that PCR effectively amplified and isolated TPO and IGF-1 genes. The enzyme digestion identification and DNA sequence analysis confirmed that the sequence is exactly the same as reported in the literature. The successful construction of the cDNA library laid the foundation for the future isolation and cloning of hepatocyte-producing cytokines, and provided an important material for studying the biological functions of hepatocytes at the molecular level.