Removing external DNA contamination from arthropod predators destined for molecular gut-content analysis
Ecological research requires large samples for statistical validity, typically hundreds or thousands of individuals, which are most efficiently gathered by mass‐collecting techniques. For the study of interspecific interactions, molecular gut‐content analysis enables detection of arthropod predation...
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Veröffentlicht in: | Molecular ecology resources 2012-05, Vol.12 (3), p.464-469 |
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description | Ecological research requires large samples for statistical validity, typically hundreds or thousands of individuals, which are most efficiently gathered by mass‐collecting techniques. For the study of interspecific interactions, molecular gut‐content analysis enables detection of arthropod predation with minimal disruption of community interactions. Field experiments have demonstrated that standard mass‐collection methods, such as sweep netting, vacuum sampling and foliage beating, sometimes lead to contamination of predators with nontarget DNA, thereby compromising resultant gut‐content data. We deliberately contaminated immature Coleomegilla maculata and Podisus maculiventris that had been fed larvae of Leptinotarsa decemlineata by topically applying homogenate of the alternate prey Leptinotarsa juncta. We then attempted to remove contaminating DNA by washing in ethanol or bleach. A 40‐min wash with end‐over‐end rotation in 80% EtOH did not reliably reduce external DNA contamination. Identical treatment with 2.5% commercial bleach removed most externally contaminating DNA without affecting the detectability of the target prey DNA in the gut. Use of this bleaching protocol, perhaps with minor modifications tailored to different predator–prey systems, should reliably eliminate external DNA contamination, thereby alleviating concerns about this possible source of cross‐contamination for mass‐collected arthropod predators destined for molecular gut‐content analysis. |
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For the study of interspecific interactions, molecular gut‐content analysis enables detection of arthropod predation with minimal disruption of community interactions. Field experiments have demonstrated that standard mass‐collection methods, such as sweep netting, vacuum sampling and foliage beating, sometimes lead to contamination of predators with nontarget DNA, thereby compromising resultant gut‐content data. We deliberately contaminated immature Coleomegilla maculata and Podisus maculiventris that had been fed larvae of Leptinotarsa decemlineata by topically applying homogenate of the alternate prey Leptinotarsa juncta. We then attempted to remove contaminating DNA by washing in ethanol or bleach. A 40‐min wash with end‐over‐end rotation in 80% EtOH did not reliably reduce external DNA contamination. Identical treatment with 2.5% commercial bleach removed most externally contaminating DNA without affecting the detectability of the target prey DNA in the gut. Use of this bleaching protocol, perhaps with minor modifications tailored to different predator–prey systems, should reliably eliminate external DNA contamination, thereby alleviating concerns about this possible source of cross‐contamination for mass‐collected arthropod predators destined for molecular gut‐content analysis.</description><identifier>ISSN: 1755-098X</identifier><identifier>EISSN: 1755-0998</identifier><identifier>DOI: 10.1111/j.1755-0998.2012.03112.x</identifier><identifier>PMID: 22268594</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Animals ; Arthropoda ; Coleomegilla maculata ; Coleoptera - physiology ; Content analysis ; Deoxyribonucleic acid ; DNA ; DNA - chemistry ; DNA - genetics ; DNA - isolation & purification ; DNA Contamination ; DNA decontamination ; Gastrointestinal Contents - chemistry ; Heteroptera - physiology ; Leptinotarsa decemlineata ; molecular gut-content analysis ; Molecular Sequence Data ; Podisus maculiventris ; Predatory Behavior ; sampling ; Sequence Analysis, DNA ; Specimen Handling - methods</subject><ispartof>Molecular ecology resources, 2012-05, Vol.12 (3), p.464-469</ispartof><rights>Published 2012. 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For the study of interspecific interactions, molecular gut‐content analysis enables detection of arthropod predation with minimal disruption of community interactions. Field experiments have demonstrated that standard mass‐collection methods, such as sweep netting, vacuum sampling and foliage beating, sometimes lead to contamination of predators with nontarget DNA, thereby compromising resultant gut‐content data. We deliberately contaminated immature Coleomegilla maculata and Podisus maculiventris that had been fed larvae of Leptinotarsa decemlineata by topically applying homogenate of the alternate prey Leptinotarsa juncta. We then attempted to remove contaminating DNA by washing in ethanol or bleach. A 40‐min wash with end‐over‐end rotation in 80% EtOH did not reliably reduce external DNA contamination. Identical treatment with 2.5% commercial bleach removed most externally contaminating DNA without affecting the detectability of the target prey DNA in the gut. Use of this bleaching protocol, perhaps with minor modifications tailored to different predator–prey systems, should reliably eliminate external DNA contamination, thereby alleviating concerns about this possible source of cross‐contamination for mass‐collected arthropod predators destined for molecular gut‐content analysis.</description><subject>Animals</subject><subject>Arthropoda</subject><subject>Coleomegilla maculata</subject><subject>Coleoptera - physiology</subject><subject>Content analysis</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>DNA - chemistry</subject><subject>DNA - genetics</subject><subject>DNA - isolation & purification</subject><subject>DNA Contamination</subject><subject>DNA decontamination</subject><subject>Gastrointestinal Contents - chemistry</subject><subject>Heteroptera - physiology</subject><subject>Leptinotarsa decemlineata</subject><subject>molecular gut-content analysis</subject><subject>Molecular Sequence Data</subject><subject>Podisus maculiventris</subject><subject>Predatory Behavior</subject><subject>sampling</subject><subject>Sequence Analysis, DNA</subject><subject>Specimen Handling - methods</subject><issn>1755-098X</issn><issn>1755-0998</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkUtv1DAUhS0Eog_4C8gSC9hk8COO7QWLqrQF1A4SAsHOumM7rYckHuwEZv49DlNmwQLhxfWV_J1zr3wQwpQsaDmv1gsqhaiI1mrBCGULwmmp2wfo-PDw8NCrr0foJOc1IQ3Rsn6MjhhjjRK6PkZ3H30ff4ThFvvt6NMAHX6zPMM2DiP0YYAxxAG3KfYY0niX4iY6vEnewRhTxs7nMQze4TYm3MfO26mDhG-nsZod_DBiKJa7HPIT9KiFLvun9_cp-nx58en8bXX94erd-dl1ZetGskqDAM1AlSJXyoJ1GnTLNXdE8Noqp6X0nAraSgBKnNDUtnblheSU87rlp-jF3neT4vep7Gf6kK3vOhh8nLLRmnOqai0L-fKfJCVEqVrWVBf0-V_oOk7zZxVK0IIIJlmh1J6yKeacfGs2KfSQdsXKzLmZtZkjMXM8Zs7N_M7NbIv02f2AadV7dxD-CaoAr_fAz9D53X8bm5uL5dwVfbXXhzz67UEP6ZtpJJfCfFleGSbl--ZGNYbzX1JYtgw</recordid><startdate>201205</startdate><enddate>201205</enddate><creator>GREENSTONE, MATTHEW H.</creator><creator>WEBER, DONALD C.</creator><creator>COUDRON, THOMAS A.</creator><creator>PAYTON, MARK E.</creator><creator>HU, JING S.</creator><general>Blackwell Publishing Ltd</general><general>Wiley Subscription Services, Inc</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SN</scope><scope>7SS</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7TM</scope><scope>7X8</scope></search><sort><creationdate>201205</creationdate><title>Removing external DNA contamination from arthropod predators destined for molecular gut-content analysis</title><author>GREENSTONE, MATTHEW H. ; WEBER, DONALD C. ; COUDRON, THOMAS A. ; PAYTON, MARK E. ; HU, JING S.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4672-9a5a92a8a927b8cacd9a9f393d0534c8d977e3151f7aa10d591cfcbe5731334f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Animals</topic><topic>Arthropoda</topic><topic>Coleomegilla maculata</topic><topic>Coleoptera - physiology</topic><topic>Content analysis</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>DNA - chemistry</topic><topic>DNA - genetics</topic><topic>DNA - isolation & purification</topic><topic>DNA Contamination</topic><topic>DNA decontamination</topic><topic>Gastrointestinal Contents - chemistry</topic><topic>Heteroptera - physiology</topic><topic>Leptinotarsa decemlineata</topic><topic>molecular gut-content analysis</topic><topic>Molecular Sequence Data</topic><topic>Podisus maculiventris</topic><topic>Predatory Behavior</topic><topic>sampling</topic><topic>Sequence Analysis, DNA</topic><topic>Specimen Handling - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>GREENSTONE, MATTHEW H.</creatorcontrib><creatorcontrib>WEBER, DONALD C.</creatorcontrib><creatorcontrib>COUDRON, THOMAS A.</creatorcontrib><creatorcontrib>PAYTON, MARK E.</creatorcontrib><creatorcontrib>HU, JING S.</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular ecology resources</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>GREENSTONE, MATTHEW H.</au><au>WEBER, DONALD C.</au><au>COUDRON, THOMAS A.</au><au>PAYTON, MARK E.</au><au>HU, JING S.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Removing external DNA contamination from arthropod predators destined for molecular gut-content analysis</atitle><jtitle>Molecular ecology resources</jtitle><addtitle>Mol Ecol Resour</addtitle><date>2012-05</date><risdate>2012</risdate><volume>12</volume><issue>3</issue><spage>464</spage><epage>469</epage><pages>464-469</pages><issn>1755-098X</issn><eissn>1755-0998</eissn><abstract>Ecological research requires large samples for statistical validity, typically hundreds or thousands of individuals, which are most efficiently gathered by mass‐collecting techniques. For the study of interspecific interactions, molecular gut‐content analysis enables detection of arthropod predation with minimal disruption of community interactions. Field experiments have demonstrated that standard mass‐collection methods, such as sweep netting, vacuum sampling and foliage beating, sometimes lead to contamination of predators with nontarget DNA, thereby compromising resultant gut‐content data. We deliberately contaminated immature Coleomegilla maculata and Podisus maculiventris that had been fed larvae of Leptinotarsa decemlineata by topically applying homogenate of the alternate prey Leptinotarsa juncta. We then attempted to remove contaminating DNA by washing in ethanol or bleach. A 40‐min wash with end‐over‐end rotation in 80% EtOH did not reliably reduce external DNA contamination. Identical treatment with 2.5% commercial bleach removed most externally contaminating DNA without affecting the detectability of the target prey DNA in the gut. Use of this bleaching protocol, perhaps with minor modifications tailored to different predator–prey systems, should reliably eliminate external DNA contamination, thereby alleviating concerns about this possible source of cross‐contamination for mass‐collected arthropod predators destined for molecular gut‐content analysis.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>22268594</pmid><doi>10.1111/j.1755-0998.2012.03112.x</doi><tpages>6</tpages></addata></record> |
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subjects | Animals Arthropoda Coleomegilla maculata Coleoptera - physiology Content analysis Deoxyribonucleic acid DNA DNA - chemistry DNA - genetics DNA - isolation & purification DNA Contamination DNA decontamination Gastrointestinal Contents - chemistry Heteroptera - physiology Leptinotarsa decemlineata molecular gut-content analysis Molecular Sequence Data Podisus maculiventris Predatory Behavior sampling Sequence Analysis, DNA Specimen Handling - methods |
title | Removing external DNA contamination from arthropod predators destined for molecular gut-content analysis |
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