Preparation of DNA suitable for PCR amplification from fresh or fixed single dinoflagellate cells.
A method is described to prepare total DNA from single cells of dinoflagellates, which can be used for PCR amplification. As model organisms, we used a stock strain of Alexandrium catenella and cells of Dinophysis acuminata harvested from the Atlantic Ocean. Fresh grown cells or cells maintained in different preservatives were tested as sources for DNA preparation. The method used to prepare DNA combines physicochemical and enzymatic procedures on cells embedded in agarose plugs or beads. The agarose pieces containing the DNA were used to perform PCR amplification of a fragment of DNA containing a 5.8S rRNA gene and the flanking internal transcribed spacers (ITS1 and ITS2).
Marín
I
I
Universidad Autónoma de Madrid Cantoblanco.
Aguilera
A
A
Reguera
B
B
Abad
J P
JP
D000818:Animals; D017931:DNA Primers; D016054:DNA, Protozoan / Q000235:genetics / Q000302:isolation & purification*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D004141:Dinoflagellida / Q000166:cytology / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012340:RNA, Ribosomal, 5.8S / Q000235:genetics
D017931:DNA Primers; D016054:DNA, Protozoan; D021903:DNA, Ribosomal Spacer; D012340:RNA, Ribosomal, 5.8S