【摘要】 将苏云金芽胞杆菌(Bacillus thuringiensis,简称Bt)8010在LB液体培养基30℃、230r/min下振荡培养,通过生长曲线测定(OD600)和显微镜观察,分别在8、30、39、42h和46h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,找到一种方法可以很有效的提取Bt3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。更多还原

【Abstract】 Bacillus thuringiensis(Bt)subsp.kurstaki 8010 was grown aerobically at 30 oC with shaking at 230 r/min 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(8h,30h,39h,42h and 46h after dilution)during the exponential phase,stationary phase and phase of lysis were recovered,immediately frozen in liquid nitrogen and stored at-70 oC until RNA extraction.An improved method was applied to extract total RNA from three growth stages of Bt 8010.Total RNA were effective extracted and the high quality RNA can be used for cDNA synthesis,RT-PCR,transcriptomics analysis and others downstream experiments.更多还原