About Zymoprep™ Yeast Plasmid Miniprep I
The Zymoprep™ 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. There is no need for glass beads, phenol, or vortexing. Instead, plasmid DNA is reliably recovered from yeast cells whether colonies, patches on plates, or liquid cultures are sampled. Plasmid yields are typically between 0.01-0.3 ng for most 2 µ based plasmids from 1.5 ml overnight cultures. This kit also works well with low copy number yeast plasmids. Recovered plasmid DNA is in TE buffer and can be used for E. coli transformation, Western blotting, PCR, etc.
|Elution Volume||≥ 35 µl|
|Plasmid DNA Size||DNA up to 23 kb.|
|Compatibility||S. cerevisiae, C. albicans and S. pombe, and other fungi species sensitive to yeast lytic enzymatic digestion (Zymolyase™).|
Use toothpick or pipette tip to pick roughly 5-15 µl volume of yeast colonies or patches from plates and dispense into 150 µl of Solution 1-enzyme mixture (add 13 µl Zymolyase™ to each ml of Solution 1 to make Solution 1-enzyme mixture).
Incubate at 37°C for 15-60 minutes (15 minutes is the minimal incubation time, although longer incubation is optional).
Add 150 µl Solution 2 to each tube. Mix well
Add 150 µl Solution 3 to each tube. Mix well.
Centrifuge at maximum speed for 2 minutes.
Transfer supernatant to new tubes. Add 400 µl isopropanol (2-propanol) to each tube. Mix well.
Centrifuge at maximum speed for 8 minutes. Aspirate supernatant. Spin briefly and remove any residual supernatant.
Resuspend the plasmid pellet in 35 µl TE buffer. It is not necessary to dry the pellet before adding TE. Sometimes the pellet needs to be pipette for complete dissolving.
Use 3-5 µl of the plasmid DNA for E. coli transformation experiments.