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目的：建立一种简便、快速检测应激诱导细胞生成的5′端转移RNA（ tRNA）半分子的方法。方法提取应激诱导细胞RNA，用poly（ A）加尾酶在RNA分子3′端加尾后，加入与待测tRNA的3′端部分序列互补的简并DNA探针杂交，经RNase H消化与DNA探针互补的tRNA序列，再由oligo（ dT） n锚定引物进行逆转录获得互补DNA（ cDNA），以5′端tRNA半分子和锚定引物的序列为PCR反应的上下游引物，PCR扩增检测5′端tRNA半分子。结果经poly（ A）加尾-RNaseH消化-RT-PCR-电泳等步骤，可以实现对5′端tRNA半分子的检测分析。结论 poly （ A）加尾-RNaseH消化-RT-PCR法的建立实现了5′端tRNA半分子的简便、快速检测分析，为tRNA半分子生物学功能研究和检测分析提供了工具。“,”Objective To develop a simple and quick method for detection of stress-induced 5′transfer RNA( tRNA) halves.Methods Total RNA purified from stress induced cells was polyadenylated by poly( A) polymerase, and then degen-erate DNA probes were used to hybridize with 3′tRNA-halves of intact tRNAs,while RNase H specifically degraded the 3′tRNA-halves strand in tRNA-DNA probes hybrids.Using the RNase H digestion total RNA as templates, complementary DNA( cDNA) was synthesized by oligo ( dT) n-anchored primers.The primer of 5′tRNA halves and anchored-primer were used to amplify 5′tRNA halves by PCR.Results The results showed that the method of poly ( A )-tailed-RNase H digestion-RT-PCR could be successfully used to detect stress-induced 5′tRNA halves.Conclusion A simple and quick method for detection of 5′tRNA halves has been established,which is a user-friendly tool for 5′tRNA halves detection and function research.