Description
The Zymoprep Yeast Plasmid Miniprep II is a simple and efficient yeast plasmid miniprep that is based on the E. coli alkaline lysis method but using Zymolyase as the first solution. The kit provides all the necessary reagents for plasmid isolation from S. cerevisiae, C. albicans and S. pombe, and any fungi whose cell walls are susceptible to yeast lytic enzyme lysis. The procedure is simple and efficient, with no need for glass beads or phenol. The Zymoprep II recovers about 5 fold more plasmid DNA compared to Zymoprep I and eliminates the isopropanol precipitation step. Reliably recover plasmid DNA from yeast colonies, patches on plates, or as liquid cultures. The system is ideal for low-copy number and hard-to-isolate plasmids. Eluted plasmid DNA can be used directly for E. Coli transformation, PCR, and Southern blot analysis.
Technical Specifications
| Applicable For |
Plasmid DNA is well suited for downstream applications such as PCR, transformation, hybridization and other sensitive applications |
| Elution Volume |
= 10 µl |
| Equipment |
Microcentrifuge & Heat Block/Bath |
| Processing Volume |
= 1.5 ml of Culture |
| Sample Source |
Cell Culture (Colonies/Patches or Liquid Culture) |
| Size Range |
Up to 23 kb. |
| Yield |
Typically between 0.01-0.3 ng for most 2 µ based plasmids from 1.5 ml overnight cultures |
Q1: What is the difference between Zymoprep Yeast Plasmid Miniprep I and Miniprep II?
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.
Q2: What is the typical plasmid yield?
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.
Q3: Can this kit be used to isolate linear plasmid DNA?
Q4: If I’m using stationary phase yeast cells, what can I do to improve sample lysis?
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.
Q5: What yeast strains are these kits compatible with?
Any strains susceptible to Zymolyase, which includes:
Susceptible fungal genera:
Asbya Kloekera
Candida Kluyveromyces
Debaryomyces Lipomyces
Eremothecium Metschikowia
Endomyces Pichia
Hansenula Pullularia
Hanseniaspora Saccharomyces
Saccaromycodes Saccharomycopsis
Schizosaccahromyces Torulopsis
