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应用套式PCR克隆ST13基因cDNA的5'端序列 被引量:2

Cloning the 5end fragment of ST13 cDNA by nested PCR
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摘要 目的获得ST13基因cDNA5′端的未知序列。方法应用套式PCR方法直接从cDNA文库中扩增该基因的5′端序列,然后将扩增的片段克隆到pGEM-T.easy载体,并进行序列分析。结果第1次PCR后得到1条约550bp的片段,经套式PCR后获得1条约480bp的片段,经序列分析证实该扩增的片段是ST13基因cDNA的5′端未知序列。结论套式PCR技术是克隆基因cDNA的5′端未知序列较简捷、有效的方法。 Objective To clone and sequence the 5end fragment of ST13 cDNA. Methods The 5end fragment of the ST13 cDNA was amplified directly from cDNA library by nested PCR method, and cloned into pGEMT.easy vector, then the sequencing of the inserted PCR product was performed. Results After primary and secondary PCR, two PCR products obtained with the size of about 550bp and 480bp, respectively, were both proved to be the 5end of ST13 cDNA by sequencing. Conclusion The method presented is very simple and effective for cloning and sequencing the 5end of a target gene.
出处 《中华医学遗传学杂志》 CAS CSCD 北大核心 1999年第3期174-176,共3页 Chinese Journal of Medical Genetics
基金 国家九五攻关项目
关键词 ST13基因 聚合酶链反应 序列 CDNA T13 geneNested polymerase chain reactionSequence
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