Rapid extraction of DNA from diverse soils by guanidine thiocyanate method.
Molecular methods are being frequently used for the study of soil microbial communities as majority of naturally occurring microbial populations are non-culturable. In the present study, we describe a protocol of DNA extraction from diverse soils using a combination of heat, enzyme (lysozyme) and guanidine thiocyanate. The efficacy of the procedure was evaluated in terms of yield, purity and duration of extraction. The protocol was effective for neutral, acidic as well as alkaline soils (pH range 4.5-8.5). The extracted soil DNA was observed with negligible shearing on agarose gel and the time taken for restriction digestion was very less. Further, the DNA extracted was almost completely devoid of contaminants and pure enough which could be used for PCR amplification and Southern hybridization.
Agarwal
A
A
Department of Microbiology, CBSH, GB Pant University of Agriculture & Technology, Pantnagar, India. nupriyaagarwal@usa.net
Kumar
C
C
Goel
R
R
D004269:DNA, Bacterial / Q000032:analysis / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006146:Guanidines / Q000737:chemistry*; D015183:Restriction Mapping; D012988:Soil Microbiology; D013861:Thiocyanates / Q000737:chemistry*
D004269:DNA, Bacterial; D006146:Guanidines; D013861:Thiocyanates; C054436:guanidine thiocyanate