Synechocystis sp.strain PCC 6803定量PCR模板制备方法的改进  

Improving methods for preparating quantitative PCR template from the cyanobacteriumSynechocystissp.strain PCC 6803

作　　者：阳桂丹 张巨源[1] 陈雯莉[1] YANG Guidan ZHANG Juyuan CHEN Wenli(State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070 , Chin)

机构地区：[1]华中农业大学农业微生物学国家重点实验室,武汉430070

出　　处：《华中农业大学学报》2016年第6期44-51,共8页Journal of Huazhong Agricultural University

基　　金：国家自然科学基金项目(31570048);中央高校基本科研业务费专项(2014PY003)

摘　　要：以Synechocystis sp.strain PCC 6803为实验菌株,设计改良制备定量PCR模板的方法;同时,对Synechocystis sp.strain PCC 6803总RNA的Trizol抽提法进行优化。结果显示,利用改良方法抽提得到的RNA结构更完整;同时,利用改良方法制备的模板可以用于后续定量PCR实验;其中,只有高表达量基因适用于半定量PCR,低表达量基因则需要实时荧光定量PCR检测。The cyanobacterium Synechocystis sp. strain PCC 6803 is one of the most important models for studying photosynthesis and ecology.Real-time fluorescent quantitative PCR and semi-quantitative PCR are extensively exploited to investigate the gene expression of this cyanobacterium. The quality of quantitative PCR template has a decisive influence on final data and the experimental results.When preparing Synechocystis sp. strain PCC 6803 RNA for quantitative PCR, degradation occurs frequently, leading to unreliable results.Methods for preparating quantitative PCR template from Synechocystis sp. strain PCC 6803 were improved.Results showed that the RNA isolated with the improved method was more intact and can be directly reverse-transcribed to the cDNA template for quantitative PCR.Using the cDNA template obtained,detecting genes with high expression can be performed simply by semi-quantitative PCR,while detection of those genes with low expression requires real-time fluorescent quantitativePCR.

关 键 词：定量PCR Synechocystissp.strain PCC6803 RNA抽提 qPCR模板制备 

分 类 号：Q93-31[生物学—微生物学]