Quantification of functional genes from procaryotes in soil by PCR.
Controlling turnover processes and fluxes in soils and other environments requires information about the gene pool and possibilities for its in situ induction. Therefore in the recent years there has been a growing interest in genes and transcripts coding for metabolic enzymes. Besides questions addressing redundancy and diversity, more and more attention is given on the abundance of specific DNA and mRNA in the different habitats. This review will describe several PCR techniques that are suitable for quantification of functional genes and transcripts such as MPN-PCR, competitive PCR and real-time PCR. The advantages and disadvantages of the mentioned methods are discussed. In addition, the problems of quantitative extraction of nucleic acid and substances that inhibit polymerase are described. Finally, some examples from recent papers are given to demonstrate the applicability and usefulness of the different approaches.
Sharma
Shilpi
S
Institute of Soil Ecology, GSF-National Research Center for Environment and Health, P.O. Box 1129, D-85764, Neuherberg, Germany.
Radl
Viviane
V
Hai
Brigitte
B
Kloos
Karin
K
Fuka
Mirna Mrkonjic
MM
Engel
Marion
M
Schauss
Kristina
K
Schloter
Michael
M
D001419:Bacteria / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D012988:Soil Microbiology
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D018076:DNA, Complementary; D012329:RNA, Bacterial