The assessment of DNA from marine organisms via a modified salting-out protocol

被引:34
作者
Ferrara, Giovanni Battista
Murgia, Barbara
Parodi, Anna Maria
Valisano, Laura
Cerrano, Carlo
Palmisano, Giulio
Bavestrello, Giorgio
Sara, Michele
机构
[1] Univ Genoa, Dipartimento Studio Territorio & Risorse, I-16132 Genoa, Italy
[2] Univ Genoa, Dipartimento Biol, I-16132 Genoa, Italy
[3] Univ Marche, Dipartimento Sci Mare, I-60131 Ancona, Italy
关键词
Symbiodinium; Porifera; Cnidaria; DNA; PCR;
D O I
10.2478/s11658-006-0013-7
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We developed a rapid, practical and non-toxic salting-out method for the extraction of DNA from marine organisms, and tested it on two representative species of Porifera and Cnidaria, both living in association with symbiotic zooxanthellae. We tested the efficiency of the protocol by comparing the output of the method for fresh tissue, frozen tissue and tissue stored in ethanol. It proved to be effective for extracting DNA in the case of the methods of preservation considered here, and for obtaining quantities of DNA comparable to those obtained via the traditional approach. The DNA from both species was of good quality. The DNA obtained was amplified by PCR using specific primers for the large ribosomal subunit, allowing the identification of the presence of both the host and symbiont genomes.
引用
收藏
页码:155 / 160
页数:6
相关论文
共 19 条
[1]  
Chen CA, 2000, MAR BIOTECHNOL, V2, P146
[2]   DNA FINGERPRINTS OF A GORGONIAN CORAL - A METHOD FOR DETECTING CLONAL STRUCTURE IN A VEGETATIVE SPECIES [J].
COFFROTH, MA ;
LASKER, HR ;
DIAMOND, ME ;
BRUENN, JA ;
BERMINGHAM, E .
MARINE BIOLOGY, 1992, 114 (02) :317-325
[3]  
COWEN R, 1998, PALAIOS, V3, P221
[4]   A novel method for the transport and analysis of genetic material from polyps and zooxanthellae of scleractinian corals [J].
Crabbe, MJC .
JOURNAL OF BIOCHEMICAL AND BIOPHYSICAL METHODS, 2003, 57 (02) :171-176
[5]   CORAL-REEF BLEACHING IN THE 1980S AND POSSIBLE CONNECTIONS WITH GLOBAL WARMING [J].
GLYNN, PW .
TRENDS IN ECOLOGY & EVOLUTION, 1991, 6 (06) :175-179
[6]   Stability of an octocoral-algal symbiosis over time and space [J].
Goulet, TL ;
Coffroth, MA .
MARINE ECOLOGY PROGRESS SERIES, 2003, 250 :117-124
[7]   How are climate and marine biological outbreaks functionally linked? [J].
Hayes, ML ;
Bonaventura, J ;
Mitchell, TP ;
Prospero, JM ;
Shinn, EA ;
Van Dolah, F ;
Barber, RT .
HYDROBIOLOGIA, 2001, 460 (1-3) :213-220
[8]   Climate change, coral bleaching and the future of the world's coral reefs [J].
Hoegh-Guldberg, O .
MARINE AND FRESHWATER RESEARCH, 1999, 50 (08) :839-866
[9]  
JUNGHANS H, 1990, BIOTECHNIQUES, V8, P176
[10]   Investigating the biodiversity, ecology, and phylogeny of endosymbiotic dinoflagellates in the genus Symbiodinium using the its region:: In search of a "species" level marker [J].
LaJeunesse, TC .
JOURNAL OF PHYCOLOGY, 2001, 37 (05) :866-880