Both the Zymoprep Yeast Plasmid Miniprep I and II utilize the same chemistry for lysis; however, Miniprep I uses isopropanol precipitation and Miniprep II utilizes a column for purification. The Miniprep II allows for consistent yield and purity; and samples can be concentrated to a low elution volume.
Typically, between 0.01 – 0.3 ng for most 2 µ based plasmid from 1.5 ml overnight cultures. In order to generate more plasmid, the plasmid is typically transformed into E. Coli , cultured, and isolated using a traditional E. Coli plasmid prep.
We generally recommend working with fresh or early log phase cells, which are easier to lyse. For stationary phase cells, user optimization is necessary, and we recommend increasing digestion to > 1 hour and/ or increasing the amount of Zymolyase.
Any strains susceptible to Zymolyase, which includes the following fungal genera:
Asbya,
Kloekera,
Candida,
Kluyveromyces,
Debaryomyces,
Lipomyces,
Eremothecium,
Metschikowia,
Endomyces,
Pichia,
Hansenula,
Pullularia,
Hanseniaspora,
Saccharomyces,
Saccaromycodes,
Saccharomycopsis,
Schizosaccahromyces,
Torulopsis