Extracting DNA from within intact foraminiferal shells
Here we present an evaluation of several methods for extracting DNA from within intact foraminiferal shells. We tested 12 lysis buffers under carefully optimised incubation conditions, evaluating their success in terms of efficiency in evacuating cellular material, crude DNA yield, PCR success, and...
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Veröffentlicht in: | Marine micropaleontology 2014-05, Vol.109, p.46-53 |
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description | Here we present an evaluation of several methods for extracting DNA from within intact foraminiferal shells. We tested 12 lysis buffers under carefully optimised incubation conditions, evaluating their success in terms of efficiency in evacuating cellular material, crude DNA yield, PCR success, and final shell integrity. A number of the buffers tested produced excellent results. The most successful method utilized a lysis buffer containing Sodium N-lauroyl sarcosine, Guanidinium isothiocyanate, Isopropanol, TRIS and NaCl, incubated at 75°C for 24h, followed by chloroform extraction and ethanol precipitation. High yields of DNA were produced, with no signs of PCR inhibition, and the foraminiferal shells were left completely intact. Retaining the shell of individual specimens presents a significant advance, allowing for direct comparisons between shell morphology and genotype data, which could greatly enhance the utility of foraminifera as palaeoproxies of past climate change.
•Methods for extracting DNA from within intact foraminiferal shells were evaluated.•12 lysis buffers were compared under optimised incubation conditions.•High yields of good quality DNA were produced for use in PCR.•The delicate shells were retained undamaged for use in morphological analyses. |
doi_str_mv | 10.1016/j.marmicro.2014.04.001 |
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•Methods for extracting DNA from within intact foraminiferal shells were evaluated.•12 lysis buffers were compared under optimised incubation conditions.•High yields of good quality DNA were produced for use in PCR.•The delicate shells were retained undamaged for use in morphological analyses.</description><identifier>ISSN: 0377-8398</identifier><identifier>EISSN: 1872-6186</identifier><identifier>DOI: 10.1016/j.marmicro.2014.04.001</identifier><language>eng</language><publisher>Elsevier B.V</publisher><subject>Cell lysis ; DNA extraction ; Foraminifera ; PCR inhibition ; Shell preservation</subject><ispartof>Marine micropaleontology, 2014-05, Vol.109, p.46-53</ispartof><rights>2014 Elsevier B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-a434t-763cfa738d6c2e048348a1f01e8d8247df9afbf726e4c47048961759fdd68cc03</citedby><cites>FETCH-LOGICAL-a434t-763cfa738d6c2e048348a1f01e8d8247df9afbf726e4c47048961759fdd68cc03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0377839814000425$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3536,27903,27904,65309</link.rule.ids></links><search><creatorcontrib>Seears, Heidi A.</creatorcontrib><creatorcontrib>Wade, Christopher M.</creatorcontrib><title>Extracting DNA from within intact foraminiferal shells</title><title>Marine micropaleontology</title><description>Here we present an evaluation of several methods for extracting DNA from within intact foraminiferal shells. We tested 12 lysis buffers under carefully optimised incubation conditions, evaluating their success in terms of efficiency in evacuating cellular material, crude DNA yield, PCR success, and final shell integrity. A number of the buffers tested produced excellent results. The most successful method utilized a lysis buffer containing Sodium N-lauroyl sarcosine, Guanidinium isothiocyanate, Isopropanol, TRIS and NaCl, incubated at 75°C for 24h, followed by chloroform extraction and ethanol precipitation. High yields of DNA were produced, with no signs of PCR inhibition, and the foraminiferal shells were left completely intact. Retaining the shell of individual specimens presents a significant advance, allowing for direct comparisons between shell morphology and genotype data, which could greatly enhance the utility of foraminifera as palaeoproxies of past climate change.
•Methods for extracting DNA from within intact foraminiferal shells were evaluated.•12 lysis buffers were compared under optimised incubation conditions.•High yields of good quality DNA were produced for use in PCR.•The delicate shells were retained undamaged for use in morphological analyses.</description><subject>Cell lysis</subject><subject>DNA extraction</subject><subject>Foraminifera</subject><subject>PCR inhibition</subject><subject>Shell preservation</subject><issn>0377-8398</issn><issn>1872-6186</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><recordid>eNqFkEtLQzEQhYMoWKt_Qe7Szb3mZZK7s9T6gKIbXYeYTGzKfdQk9fHvTamuhQOzmHMOMx9C5wQ3BBNxuW56E_tg49hQTHiDizA5QBOiJK0FUeIQTTCTslasVcfoJKU1Lg7BxASJxVeOxuYwvFU3j7PKx7GvPkNehaEKQy6byo_R9GEIHqLpqrSCrkun6MibLsHZ75yil9vF8_y-Xj7dPcxny9pwxnMtBbPeSKacsBQwV4wrQzwmoJyiXDrfGv_qJRXALZfF0Aoir1rvnFDWYjZFF_veTRzft5Cy7kOy5QIzwLhNmggqW4EVlcUq9tYCIqUIXm9iKGS-NcF6B0qv9R8ovQOlcREmJXi9D0J55CNA1MkGGCy4EMFm7cbwX8UPyf10fw</recordid><startdate>20140501</startdate><enddate>20140501</enddate><creator>Seears, Heidi A.</creator><creator>Wade, Christopher M.</creator><general>Elsevier B.V</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7TM</scope><scope>7TN</scope><scope>F1W</scope><scope>H95</scope><scope>L.G</scope></search><sort><creationdate>20140501</creationdate><title>Extracting DNA from within intact foraminiferal shells</title><author>Seears, Heidi A. ; Wade, Christopher M.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a434t-763cfa738d6c2e048348a1f01e8d8247df9afbf726e4c47048961759fdd68cc03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Cell lysis</topic><topic>DNA extraction</topic><topic>Foraminifera</topic><topic>PCR inhibition</topic><topic>Shell preservation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Seears, Heidi A.</creatorcontrib><creatorcontrib>Wade, Christopher M.</creatorcontrib><collection>CrossRef</collection><collection>Nucleic Acids Abstracts</collection><collection>Oceanic Abstracts</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><jtitle>Marine micropaleontology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Seears, Heidi A.</au><au>Wade, Christopher M.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Extracting DNA from within intact foraminiferal shells</atitle><jtitle>Marine micropaleontology</jtitle><date>2014-05-01</date><risdate>2014</risdate><volume>109</volume><spage>46</spage><epage>53</epage><pages>46-53</pages><issn>0377-8398</issn><eissn>1872-6186</eissn><abstract>Here we present an evaluation of several methods for extracting DNA from within intact foraminiferal shells. We tested 12 lysis buffers under carefully optimised incubation conditions, evaluating their success in terms of efficiency in evacuating cellular material, crude DNA yield, PCR success, and final shell integrity. A number of the buffers tested produced excellent results. The most successful method utilized a lysis buffer containing Sodium N-lauroyl sarcosine, Guanidinium isothiocyanate, Isopropanol, TRIS and NaCl, incubated at 75°C for 24h, followed by chloroform extraction and ethanol precipitation. High yields of DNA were produced, with no signs of PCR inhibition, and the foraminiferal shells were left completely intact. Retaining the shell of individual specimens presents a significant advance, allowing for direct comparisons between shell morphology and genotype data, which could greatly enhance the utility of foraminifera as palaeoproxies of past climate change.
•Methods for extracting DNA from within intact foraminiferal shells were evaluated.•12 lysis buffers were compared under optimised incubation conditions.•High yields of good quality DNA were produced for use in PCR.•The delicate shells were retained undamaged for use in morphological analyses.</abstract><pub>Elsevier B.V</pub><doi>10.1016/j.marmicro.2014.04.001</doi><tpages>8</tpages></addata></record> |
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subjects | Cell lysis DNA extraction Foraminifera PCR inhibition Shell preservation |
title | Extracting DNA from within intact foraminiferal shells |
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