An anion-exchange method to concentrate dissolved DNA from aquifer water

作者：Li Z^*; Briggs B R; Sheridan P P; Shields M S

来源：Journal of Microbiological Methods, 2013, 93(1): 1-8.

DOI：10.1016/j.mimet.2013.01.014

• 16S rRNA
• Anion-exchange
• Aquifer
• Dissolved DNA
• PCR

A rapid DNA isolation method was developed to concentrate dissolved DNA (dDNA) in aquifer water for molecular analysis. The aquifer dDNA from the Eastern Snake River Plain Aquifer (ESRPA) was extracted and concentrated using a new method with an anion-exchange Mustang® Q membrane. The concentration of aquifer dDNA in this study ranged from 60 to 264.5ngl-1 in ESRPA aquifer wells. DNA stability in ESRPA aquifer water was also tested in this study. The dDNA extracted from aquifer water samples was used for PCR amplification of bacterial 16S rRNA genes for terminal restriction fragment length polymorphism (T-RFLP) analysis and construction of 16S rRNA gene clone libraries. The ureC gene, IncP, IncQ and IncW plasmid genes were also PCR amplified from dDNA samples. Based on the results, dDNA is relatively stable in aquifer water and can be concentrated by Q membrane method for molecular analysis. The quality of isolated dDNA was suitable as a PCR template.

• 单位
  Miami University, Oxford