Table 2.

Comparison of methods of extracting DNA from diluted cultures of C. albicans

MethodReference or sourceSensitivity (cells/ml)aTime (h)Ease of use and recovery
Zymolyase, SDS, proteinase K, phenolBuchman et al. (4)10 (1) or inhibition (1)2Handy, phenol, two enzymatic steps, good recovery or total inhibition
Zymolyase, proteinase K, phenol, RNaseHolm et al. (9)10 (1) and 102(1)4Tedious, phenol, three enzymatic steps, good recovery
Glass beads, phenolSanglard et al. (19)102 (2)0.75Handy, phenol, medium recovery
Zymolyase, proteinase K, lysing solution, RNaseSanglard et al. (20)106(2)2Handy, no phenol, three enzymatic steps, low recovery
Zymolyase, silica beadsBoom et al. (3)102 (2)2.5Handy, no phenol, one enzymatic step, medium recovery, some inhibition
Zymolyase, proteinase K, silica membraneQiagen102(1)2Very handy, no phenol, two enzymatic steps, medium recovery
Zymolyase, proteinase K, silica membraneMacherey & Nagel10 (3)2Very handy, no phenol, two enzymatic steps, good recovery
Proteinase K, silica membraneb Qiagen (modified protocol)10 (23)1.5Simple, rapid, no phenol, one enzymatic step, good recovery
  • a Tenfold serial dilutions of an overnight C. albicans culture in water were obtained as described in Materials and Methods, and cell quantification was performed as explained in Materials and Methods. A 10-μl aliquot of the DNA extracted from each dilution was used for PCR amplification. A 10-μl aliquot of the PCR product was used for detection by agarose gel electrophoresis with EtBr staining. The sensitivity was defined as the highest dilution which produced a visible band by EtBr staining after agarose gel electrophoresis with the SAP primers and the PCR protocol described in the text. The number of separate experiments is indicated in parentheses.

  • b This method was adopted for the present study.