MasterPure™ Yeast DNA Purification Kit

Poster on rapid fungal DNA typing from ASM 2006 (2.3 MB)

Applications

• Purification of DNA from yeast for many molecular biology applications, including PCR amplification, restriction endonuclease digestion, Southern blotting, and genomic library preparation.
• Identification and typing of fungi.

The MasterPure™ Yeast DNA Purification Kit enables efficient, high-yield purification of high-molecular-weight DNA from yeast and other fungi. The protocol involves nonenzymatic cell lysis at 65°C, followed by removal of protein by precipitation, and nucleic acid precipitation and resuspension. No lyticase, proteolytic enzymes, or bead-beating are used in the procedure. Yeast genomic DNA yields using the MasterPure Kit are much higher than yields obtained with other commercially available kits (Fig. 1).¹ The protocol can be easily adjusted for larger or smaller samples, including single yeast colonies.¹ The recovered nucleic acid can be used directly in most applications, including PCR amplification.

Benefits

• Higher yields of yeast chromosomal DNA than with other kits.
• Recover DNA from a wide variety of yeast species, including Candida, Saccharomyces, Pichia, and Schizosaccharomyces, and filamentous fungi such as Aspergillus² and Penicillium.
• Purify DNA in less than 40 minutes.
• No enzymatic lysis, columns, or phenol or other organic extractions. No bead-beating.
• The recovered DNA has a high molecular weight (Fig. 2), and is suitable for many applications, including PCR amplification.

References

1. Hoffman, L. and Moan, E. (1998) EPICENTRE Forum 5(4), 1.
2. Jin, J., Lee, Y.-K. and Wickes, B.L. (2004) J. Clin Microbiol. 42, 4293.

Figure 1. Comparative yields of DNA from the MasterPure™ Yeast DNA Purification Kit and competitor's kits. DNA was quantitated specifically with Hoechst fluorescent dye 33258, which gives minimal fluorescence with RNA. The data represent the average DNA yields determined by fluorometry from two experiments with S. cerevisiae and C. albicans. The MasterPure Kit produced up to 17 times more DNA from C. albicans and 12 times more DNA from S. cerevisiae than other kits.