Comparison of different protocols for the extraction of microbial DNA from reef corals

This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.

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Main Authors: Santos,H.F., Carmo,F.L., Leite,D.C.A., Jesus,H.E., De Carvalho Maalouf,P., Almeida,C., Soriano,A.U., Altomari,D., Suhett,L., Vólaro,V., Valoni,E., Francisco,M., Vieira,J., Rocha,R., Sardinha,B.L., Mendes,L.B., João,R.R., Lacava,B., Jesus,R.F., Sebastian,G.V., Pessoa,A., van Elsas,J.D., Rezende,R.P., Pires,D.O., Duarte,G., Castro,C.B., Rosado,A.S., Peixoto,R.S.
Format: Digital revista
Language:English
Published: Sociedade Brasileira de Microbiologia 2012
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822012000200012
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spelling oai:scielo:S1517-838220120002000122012-08-07Comparison of different protocols for the extraction of microbial DNA from reef coralsSantos,H.F.Carmo,F.L.Leite,D.C.A.Jesus,H.E.De Carvalho Maalouf,P.Almeida,C.Soriano,A.U.Altomari,D.Suhett,L.Vólaro,V.Valoni,E.Francisco,M.Vieira,J.Rocha,R.Sardinha,B.L.Mendes,L.B.João,R.R.Lacava,B.Jesus,R.F.Sebastian,G.V.Pessoa,A.van Elsas,J.D.Rezende,R.P.Pires,D.O.Duarte,G.Castro,C.B.Rosado,A.S.Peixoto,R.S. coral microbial diversity DNA extraction This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.info:eu-repo/semantics/openAccessSociedade Brasileira de MicrobiologiaBrazilian Journal of Microbiology v.43 n.2 20122012-06-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822012000200012en10.1590/S1517-83822012000200012
institution SCIELO
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countrycode BR
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region America del Sur
libraryname SciELO
language English
format Digital
author Santos,H.F.
Carmo,F.L.
Leite,D.C.A.
Jesus,H.E.
De Carvalho Maalouf,P.
Almeida,C.
Soriano,A.U.
Altomari,D.
Suhett,L.
Vólaro,V.
Valoni,E.
Francisco,M.
Vieira,J.
Rocha,R.
Sardinha,B.L.
Mendes,L.B.
João,R.R.
Lacava,B.
Jesus,R.F.
Sebastian,G.V.
Pessoa,A.
van Elsas,J.D.
Rezende,R.P.
Pires,D.O.
Duarte,G.
Castro,C.B.
Rosado,A.S.
Peixoto,R.S.
spellingShingle Santos,H.F.
Carmo,F.L.
Leite,D.C.A.
Jesus,H.E.
De Carvalho Maalouf,P.
Almeida,C.
Soriano,A.U.
Altomari,D.
Suhett,L.
Vólaro,V.
Valoni,E.
Francisco,M.
Vieira,J.
Rocha,R.
Sardinha,B.L.
Mendes,L.B.
João,R.R.
Lacava,B.
Jesus,R.F.
Sebastian,G.V.
Pessoa,A.
van Elsas,J.D.
Rezende,R.P.
Pires,D.O.
Duarte,G.
Castro,C.B.
Rosado,A.S.
Peixoto,R.S.
Comparison of different protocols for the extraction of microbial DNA from reef corals
author_facet Santos,H.F.
Carmo,F.L.
Leite,D.C.A.
Jesus,H.E.
De Carvalho Maalouf,P.
Almeida,C.
Soriano,A.U.
Altomari,D.
Suhett,L.
Vólaro,V.
Valoni,E.
Francisco,M.
Vieira,J.
Rocha,R.
Sardinha,B.L.
Mendes,L.B.
João,R.R.
Lacava,B.
Jesus,R.F.
Sebastian,G.V.
Pessoa,A.
van Elsas,J.D.
Rezende,R.P.
Pires,D.O.
Duarte,G.
Castro,C.B.
Rosado,A.S.
Peixoto,R.S.
author_sort Santos,H.F.
title Comparison of different protocols for the extraction of microbial DNA from reef corals
title_short Comparison of different protocols for the extraction of microbial DNA from reef corals
title_full Comparison of different protocols for the extraction of microbial DNA from reef corals
title_fullStr Comparison of different protocols for the extraction of microbial DNA from reef corals
title_full_unstemmed Comparison of different protocols for the extraction of microbial DNA from reef corals
title_sort comparison of different protocols for the extraction of microbial dna from reef corals
description This study aimed to test different protocols for the extraction of microbial DNA from the coral Mussismilia harttii. Four different commercial kits were tested, three of them based on methods for DNA extraction from soil (FastDNA SPIN Kit for soil, MP Bio, PowerSoil DNA Isolation Kit, MoBio, and ZR Soil Microbe DNA Kit, Zymo Research) and one kit for DNA extraction from plants (UltraClean Plant DNA Isolation Kit, MoBio). Five polyps of the same colony of M. harttii were macerated and aliquots were submitted to DNA extraction by the different kits. After extraction, the DNA was quantified and PCR-DGGE was used to study the molecular fingerprint of Bacteria and Eukarya. Among the four kits tested, the ZR Soil Microbe DNA Kit was the most efficient with respect to the amount of DNA extracted, yielding about three times more DNA than the other kits. Also, we observed a higher number and intensities of DGGE bands for both Bacteria and Eukarya with the same kit. Considering these results, we suggested that the ZR Soil Microbe DNA Kit is the best adapted for the study of the microbial communities of corals.
publisher Sociedade Brasileira de Microbiologia
publishDate 2012
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822012000200012
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