苏云金芽胞杆菌不同分化发育时期总RNA的有效分离
Effective Isolation of Total RNA from Bacillus thuringiensis
将Bt菌株8010在LB液体培养基30℃、230rpm下振荡培养，通过生长曲线测定（OD600）和显微镜观察，分别在8 h、30 h、39 h、42 h和46 h取样得到营养生长期、孢子囊期和裂解期样品来提取总RNA。通过对试剂提取方法的改进，我们找到一种方法可以很有效的提取Bt 3个不同分化发育时期（营养生长期、孢子囊期和裂解期）的总RNA。提取的总RNA质量很好，可以进行cDNA合成、RT-PCR和转录组学研究等下游实验。
Bt subsp. kurstaki 8010 was grown aerobically at 30 ìC with shaking at 230 rpm in Luria-Bertaini (LB) medium. To obtain samples from different growth stages of Bt 8010, the growth of cells was monitored by measurements of optical density at 600 nm periodically with a spectrophotometer (Pharmacia, Sweden) and a microscrope. Cells obtained at 5 different time points (8 h, 30 h, 39 h, 42 h and 46 h after dilution) during the exponential phase, stationary phase and phase of lysis were recovered, immediately frozen in liquid nitrogen and stored at -70 ìC until RNA extraction. Total RNA was extracted using SV Total RNA Isolation System (Promega, USA) with minor modifications. An improved method was applied to extract total RNA from three growth stages of Bacillus thuringiensis. Total RNA were effective extracted and the high quality RNA can be used for cDNA synthesis, RT-PCR, transcriptomics analysis and others downstream experiment．
黄志鹏、黄天培、潘洁茹、关雄、洪永聪、李今煜
分子生物学微生物学
苏云金芽孢杆菌总RNA提取RT-PCR转录组学
Bacillus thuringiensis Total RNA Extraction RT-PCR Transcriptomics
黄志鹏,黄天培,潘洁茹,关雄,洪永聪,李今煜.苏云金芽胞杆菌不同分化发育时期总RNA的有效分离[EB/OL].(2008-01-15)[2025-08-02].http://www.paper.edu.cn/releasepaper/content/200801-452.点此复制
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