Nucleic acid isolation from environmental aqueous samples.
The application of molecular techniques has revolutionized freshwater and marine ecology, especially for plankton research. Methods, such as denatured gradient gel electrophoresis (DGGE), temperature gradient gel electrophoresis (TGGE), and single-strand conformation polymorphism (SSCP), together with environmental clone libraries, have unraveled an unexpected biodiversity of organisms in the water column. Molecular probes are just entering the field of commercialization for monitoring toxic algal blooms. Genomics and metagenomics were recently introduced into marine biology. At the basis of all molecular approaches is the isolation of nucleic acids from cultures, tissue, or environmental samples. Here, we summarize methods, quality controls, and hints for sample treatment to reliably isolate nucleic acids, both DNA and RNA, from environmental aqueous samples. This chapter not only is directed to researchers inexperienced with such methods but also is an aid to those already working in the field. It may be used as a step-by-step guide for nucleic acid isolation from field samples, and we make suggestions for subsequent use of the DNA/RNA.
Valentin
Klaus
K
Alfred Wegener Institute, D-27570 Bremerhaven, Germany.
John
Uwe
U
Medlin
Linda
L
D044822:Biodiversity; D011232:Chemical Precipitation; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004262:DNA Restriction Enzymes; D017753:Ecosystem; D005618:Fresh Water; D008386:Marine Biology; D008722:Methods; D009693:Nucleic Acid Hybridization; D009696:Nucleic Acids / Q000235:genetics / Q000302:isolation & purification*; D010933:Plankton / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011786:Quality Control; D012313:RNA / Q000235:genetics / Q000302:isolation & purification
D009696:Nucleic Acids; D012313:RNA; D004247:DNA; D004262:DNA Restriction Enzymes