利用升华法估算天然样品中细菌细胞丰度的新方法。
New method for estimating bacterial cell abundances in natural samples by use of sublimation.
作者信息
Glavin Daniel P, Cleaves H James, Schubert Michael, Aubrey Andrew, Bada Jeffrey L
机构信息
NASA Goddard Space Flight Center, Greenbelt, Maryland 20071, USA.
出版信息
Appl Environ Microbiol. 2004 Oct;70(10):5923-8. doi: 10.1128/AEM.70.10.5923-5928.2004.
Abstract
We have developed a new method based on the sublimation of adenine from Escherichia coli to estimate bacterial cell counts in natural samples. To demonstrate this technique, several types of natural samples, including beach sand, seawater, deep-sea sediment, and two soil samples from the Atacama Desert, were heated to a temperature of 500 degrees C for several seconds under reduced pressure. The sublimate was collected on a cold finger, and the amount of adenine released from the samples was then determined by high-performance liquid chromatography with UV absorbance detection. Based on the total amount of adenine recovered from DNA and RNA in these samples, we estimated bacterial cell counts ranging from approximately 10(5) to 10(9) E. coli cell equivalents per gram. For most of these samples, the sublimation-based cell counts were in agreement with total bacterial counts obtained by traditional DAPI (4,6-diamidino-2-phenylindole) staining.
摘要
我们开发了一种基于腺嘌呤从大肠杆菌升华的新方法，用于估计自然样品中的细菌细胞数量。为了演示该技术，将几种类型的自然样品，包括沙滩沙、海水、深海沉积物以及来自阿塔卡马沙漠的两个土壤样品，在减压下加热至500摄氏度几秒钟。升华物收集在冷阱上，然后通过具有紫外吸收检测的高效液相色谱法测定样品中释放的腺嘌呤量。根据从这些样品中的DNA和RNA回收的腺嘌呤总量，我们估计细菌细胞数量范围为每克约10(5)至10(9)个大肠杆菌细胞当量。对于这些样品中的大多数，基于升华的细胞计数与通过传统的DAPI（4,6-二脒基-2-苯基吲哚）染色获得的总细菌计数一致。
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