A simple method of DNA extraction from soil for detection of composite transgenic plants by PCR

Molecular Investigations of the Soil, Rhizosphere and Transgenic Glufosinate-Resistant Rape and Maize Plants in Combination with Herbicide (Basta®) Application under Field Conditions

“…From soil samples DNA was exactly isolated according to Ernst et al (1996). This procedure resulted in the isolation of DNA suitable for PCR analysis and eliminating interfering substances.…”

“…PCR amplification of DNA in soil samples can be inhibited by co-extracted humic acids. However, addition of CaCl 2 (Ernst et al, 1996) or NH 4 Al(SO 4 ) 2 (Lerat et al, 2005) to the DNA extraction buffer resulted in a successful amplification of transgenic DNA sequences. pat DNA sequences could be verified by PCR for up to 4 weeks after ploughing chopped rape plants into the soil (Fig.…”

“…Compost and litter are special microcosms harbouring soil microorganisms with high diversity. It has been shown that transgenic DNA can be detected in compost for a period of 24 months (Ernst et al, 1996(Ernst et al, , 1998Sandermann et al, 1997;Gebhard and Smalla, 1999). In addition to other niches, compost, with its highly active microbial degradation community, represents a possible hot spot for HGT.…”

Soil pretreatment and fast cell lysis for direct polymerase chain reaction from forest soils for terminal restriction fragment length polymorphism analysis of fungal communities

“…Prewashing of soil with solutions such as 50 mM Tris, 20 mM ethylenediaminetetraacetic acid (EDTA), 100 mM NaCl, and 1% PVPP (hereinafter referred to as TENP) or phosphate-buffered saline (PBS) improves DNA purity, 9 , 10 , 11 but trace amounts of humic substances unavoidably remain in DNA samples. Multivalent cations (Ca 2+ and Al 3+ ) can be used to precipitate humic substances by chemical flocculation, 12 , 13 , 14 , 15 however, it is difficult to control the concentration of the cations, and this method can also cause DNA coprecipitation. 13 , 14 Therefore, neither prewashing nor chemical flocculation alone leads to the best performance.…”

“…Multivalent cations (Ca 2+ and Al 3+ ) can be used to precipitate humic substances by chemical flocculation, 12 , 13 , 14 , 15 however, it is difficult to control the concentration of the cations, and this method can also cause DNA coprecipitation. 13 , 14 Therefore, neither prewashing nor chemical flocculation alone leads to the best performance.…”