Stable isotope probing to study functional components of complex microbial ecosystems.

Stable isotope probing to study functional components of complex microbial ecosystems.
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稳定同位素探测研究复杂微生物生态系统的功能成分。

DOI:
10.1007/978-1-62703-712-9_14
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发表时间:
2014
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Mazard S
Mazard S
中科院分区:
--
文献类型:
--
作者:
Mazard S

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本协议提出了一种方法,解剖DNA或RNA的关键生物参与一个特定的生化过程中,在一个复杂的生态系统。稳定同位素探测(SIP)允许标记和分离参与重要生化转化的群落成员的核酸,但通常不是一个群落中数量最多的成员。这种完全不依赖培养的技术规避了传统微生物分离技术或大规模全群落宏基因组研究数据挖掘的局限性,以梳理参与生物营养循环的微生物的身份和基因组库。只要有合适的稳定同位素基质,SIP实验几乎可以应用于任何正在研究的生态系统和生化途径。这种通用的方法允许进行广泛的分析,从脂肪酸分析,群落结构和生态学研究,以及涉及核酸测序的靶向宏基因组学。SIP实验通过专门针对感兴趣的生物体或生化转化,为大规模的全群落宏基因组研究提供了有效的替代方案,从而减少了测序工作和耗时的大型数据集生物信息学分析。
This protocol presents a method of dissecting the DNA or RNA of key organisms involved in a specific biochemical process within a complex ecosystem. Stable isotope probing (SIP) allows the labelling and separation of nucleic acids from community members that are involved in important biochemical transformations, yet are often not the most numerically abundant members of a community. This pure culture-independent technique circumvents limitations of traditional microbial isolation techniques or data mining from large-scale whole-community metagenomic studies to tease out the identities and genomic repertoires of microorganisms participating in biological nutrient cycles. SIP experiments can be applied to virtually any ecosystem and biochemical pathway under investigation provided a suitable stable isotope substrate is available. This versatile methodology allows a wide range of analyses to be performed, from fatty-acid analyses, community structure and ecology studies, and targeted metagenomics involving nucleic acid sequencing. SIP experiments provide an effective alternative to large-scale whole-community metagenomic studies by specifically targeting the organisms or biochemical transformations of interest, thereby reducing the sequencing effort and time-consuming bioinformatics analyses of large datasets.
同位素富集 DNA 的宏基因组分析。
DOI: 10.1007/978-1-60761-823-2_4
发表时间: 2010
期刊: Methods in molecular biology (Clifton, N.J.)
影响因子: --
作者:
Chen Y
通讯作者: Chen Y
通过 15N 稳定同位素探测鉴定六氢-1,3,5-三硝基-1,3,5-三嗪降解微生物。
DOI: --
发表时间: 2009
影响因子: 11.4
作者:
H. Roh;ChangJiang Yu;M. Fuller;K. Chu
通讯作者: K. Chu
DOI: 10.1038/ismej.2008.64
发表时间: 2008-11-01
期刊: ISME JOURNAL
影响因子: 11
作者:
Jehmlich, Nico;Schmidt, Frank;Vogt, Carsten
通讯作者: Vogt, Carsten