Isolation of marine meiofauna from sandy sediments From decanting to DNA extraction

Fonseca, V. G.; Packer, Margaret; Carvalho, Gary; Power, Deborah; Lambshead, John; Creer, S.

http://hdl.handle.net/10400.1/5414

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Abstract(s)

This protocol describes the separation of marine meiofauna from sediment and subsequent environmental DNA extraction. In this study meiofauna samples were taken with a 45 mm core from the upper 5 to 10 cm of sediment layer. Separation from sediment was achieved using a decantation process followed by isolation from fine silt using repetitive centrifugation steps with a 1.16 specific gravity (sg) LUDOX-TM solution. Meiofauna were deliberately separated from macrofauna by using a 1 mm sieve on top of a bottle-top sterile 45 !M sieve. High quality DNA was subsequently obtained using the QIAamp DNA Blood Maxi Kit (Qiagen) with minor adjustments to the manufacturer’s protocol. This procedure allowed efficient isolation of meiofaunal representatives from marine sediments and also extraction of high quality environmental DNA that can be used for downstream metagenetic analysis.

Keywords

Marine sediments Meiofauna Decanting LUDOX-TM DNA extraction

URI

http://hdl.handle.net/10400.1/5414

Citation

Vera Fonseca , Margaret Packer , Gary Carvalho , Deborah Power , John Lambshead & Simon Creer, "Isolation of marine meiofauna from sandy sediments From decanting to DNA extraction" in Protocol Exchange (2011).