| FISH技术定量解析亚硝酸盐氧化菌的条件优化 |
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| 引用本文： | 张星,林炜铁,朱雅楠.FISH技术定量解析亚硝酸盐氧化菌的条件优化[J].环境科学学报,2009,29(4):716-722. |
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| 作者姓名： | 张星 林炜铁 朱雅楠 |
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| 作者单位： | 华南理工大学生物科学与工程学院,广州,510006 |
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| 基金项目： | 广东省农业科技攻关计划 |
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| 摘 要： | 利用基于亚硝酸盐氧化菌(nitrite-oxidizing bacteria,NOB)16S rRNA序列的荧光探针,优化了针对亚硝酸盐氧化菌的FISH技术实验条件.确定了FISH技术定量检测亚硝酸盐氧化菌的具体方法,并优化了样品预处理的实验条件为热固定2h,4%多聚甲醛固定15min,乙醇脱水5min;杂交过程的实验条件为杂交温度46℃,杂交时间3h,洗脱液中NaCl浓度为60mmol·L-1.运用建立的FISH技术检测了7d内亚硝酸盐氧化菌菌群的丰度变化,充分显示了荧光原位杂交技术(Fluorescence in Situ Hybridization,FISH)的快速定性定量检测优势.
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| 关 键 词： | 荧光原位杂交 亚硝酸盐氧化菌 优化 |
| 收稿时间： | 7/3/2008 12:00:00 AM |
| 修稿时间： | 2008/9/22 0:00:00 |
Optimization of FISH to detect nitrite-oxidizing bacteria quantitatively |
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| ZHANG Xing,LIN Weitie and ZHU Yanan.Optimization of FISH to detect nitrite-oxidizing bacteria quantitatively[J].Acta Scientiae Circumstantiae,2009,29(4):716-722. |
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| Authors: | ZHANG Xing LIN Weitie and ZHU Yanan |
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| Institution: | ZHANG Xing,LIN Weitie,ZHU Yanan College of bioscience , bioengineering,South China University of Technology,Guangzhou 510006 |
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| Abstract: | Experimental conditions of Fluorescence in Situ Hybridization(FISH)for detecting nitrite-oxidizing bacteria(NOB)are optimized through orthogonal tests,by using the common fluorescent probe based on the 16 S rRNA sequence of nitrite-oxidizing bacteria.Our results showed that the optimal conditions for sample preparation were a heat fixing-time of 2 hours,4% paraformaldehyde fixing-time of 15 minutes,and ethanol dehydration time of 5 minutes.The optimal conditions for detecting NOB consisted of a hybridizatio... |
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| Keywords: | fluorescence in situ hybridization nitrite-oxidizing bacteria optimization |
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