skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: SIPSim: A Modeling Toolkit to Predict Accuracy and Aid Design of DNA-SIP Experiments

Journal Article · · Frontiers in Microbiology

DNA Stable isotope probing (DNA-SIP) is a powerful method that links identity to function within microbial communities. The combination of DNA-SIP with multiplexed high throughput DNA sequencing enables simultaneous mapping of in situ assimilation dynamics for thousands of microbial taxonomic units. Hence, high throughput sequencing enabled SIP has enormous potential to reveal patterns of carbon and nitrogen exchange within microbial food webs. There are several different methods for analyzing DNA-SIP data and despite the power of SIP experiments, it remains difficult to comprehensively evaluate method accuracy across a wide range of experimental parameters. We have developed a toolset (SIPSim) that simulates DNA-SIP data, and we use this toolset to systematically evaluate different methods for analyzing DNA-SIP data. Specifically, we employ SIPSim to evaluate the effects that key experimental parameters (e.g., level of isotopic enrichment, number of labeled taxa, relative abundance of labeled taxa, community richness, community evenness, and beta-diversity) have on the specificity, sensitivity, and balanced accuracy (defined as the product of specificity and sensitivity) of DNA-SIP analyses. Furthermore, SIPSim can predict analytical accuracy and power as a function of experimental design and community characteristics, and thus should be of great use in the design and interpretation of DNA-SIP experiments.

Research Organization:
Cornell Univ., Ithaca, NY (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division
Grant/Contract Number:
SC0004486; SC0010558; SC0016364
OSTI ID:
1510497
Alternate ID(s):
OSTI ID: 1864327
Journal Information:
Frontiers in Microbiology, Vol. 9; ISSN 1664-302X
Publisher:
Frontiers Research FoundationCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 29 works
Citation information provided by
Web of Science

References (32)

Equilibrium Sedimentation of Macromolecules in Density Gradients journal July 1957
Non-cyanobacterial diazotrophs mediate dinitrogen fixation in biological soil crusts during early crust formation journal June 2015
Using analytical ultracentrifugation to study compositional variation in vertebrate genomes journal August 2003
DNA buoyant density shifts during 15N-DNA stable isotope probing journal September 2007
Intra-genomic variation in G + C content and its implications for DNA stable isotope probing: DNA-SIP and intra-genomic variation in G + C content journal October 2014
Biphenyl-utilizing bacteria and their functional genes in a pine root zone contaminated with polychlorinated biphenyls (PCBs) journal May 2007
MFEprimer: multiple factor evaluation of the specificity of PCR primers journal November 2008
GenBank journal November 2009
Sedimentation equilibrium of DNA samples heterogeneous in density journal September 1972
DNA isolation from soil samples for cloning in different hosts collection January 2004
Determination of the base composition of deoxyribonucleic acid from its buoyant density in CsCl journal June 1962
Use of Inorganic and Organic Nitrogen by Synechococcus spp. and Diatoms on the West Florida Shelf as Measured Using Stable Isotope Probing journal September 2009
Stable Isotope Probing with 15N Achieved by Disentangling the Effects of Genome G+C Content and Isotope Enrichment on DNA Density journal March 2007
Differential abundance analysis for microbial marker-gene surveys journal September 2013
Waste Not, Want Not: Why Rarefying Microbiome Data Is Inadmissible journal April 2014
Microbial seed banks: the ecological and evolutionary implications of dormancy journal January 2011
DNA stable-isotope probing journal April 2007
Enhanced sensitivity of DNA- and rRNA-based stable isotope probing by fractionation and quantitative analysis of isopycnic centrifugation gradients: Quantitative analysis of SIP gradients journal December 2003
DNA-based stable isotope probing: a link between community structure and function journal June 2009
Importance of DNA quality in comparative soil microbial community structure analyses journal June 2008
Density-Gradient centrifugation of DNA in a fixed-angle rotor journal November 1966
DNA isolation from soil samples for cloning in different hosts journal June 2004
Datura genome reveals duplications of psychoactive alkaloid biosynthetic genes and high mutation rate following tissue culture journal March 2021
Unearthing the Ecology of Soil Microorganisms Using a High Resolution DNA-SIP Approach to Explore Cellulose and Xylose Metabolism in Soil journal May 2016
Quantitative Microbial Ecology through Stable Isotope Probing journal August 2015
Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2 journal December 2014
Waste Not, Want Not: Why Rarefying Microbiome Data is Inadmissible text January 2013
Density gradient centrifugation in angle-head rotors journal December 1964
Niche partitioning of the ubiquitous and ecologically relevant NS5 marine group journal February 2022
High-Sensitivity Stable-Isotope Probing by a Quantitative Terminal Restriction Fragment Length Polymorphism Protocol journal October 2011
Bias caused by template annealing in the amplification of mixtures of 16S rRNA genes by PCR. journal January 1996
Comparative Study of Methods for Extraction and Purification of Environmental DNA From Soil and Sludge Samples journal January 2006

Cited By (6)

Simulating metagenomic stable isotope probing datasets with MetaSIPSim journal January 2020
Evidence for phylogenetically and catabolically diverse active diazotrophs in deep-sea sediment journal January 2020
Stable isotope probing and metagenomics highlight the effect of plants on uncultured phenanthrene-degrading bacterial consortium in polluted soil journal March 2019
MFEprimer-3.0: quality control for PCR primers journal May 2019
Divergent microbial communities in groundwater and overlying soils exhibit functional redundancy for plant-polysaccharide degradation journal March 2019
Ancylomarina psychrotolerans sp. nov., isolated from sediments of Fildes Peninsula and emended the description of genus Ancylomarina journal February 2018