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