Frozen-EZ Yeast Transformation II Kit
- Fast: Yeast cells with high transformation efficiencies can be prepared in under 10 minutes.
- Simple: Easy method to transform with single or multiple plasmids in ≤ 1 hour with carrier DNA.
- Versatile: Can be used with S. cerevisiae, as well as other fungi, including C. albicans, S. pombe, and P. pastoris. Compatible with both circular and linear DNA.
|Competent Cell Stability||≥ 1 year at -70°C|
|Processing Time||≤ 15 minutes|
|Product Storage||0 - 4°C|
|Transformation DNA Input||0.2 - 1.0 µg|
|Transformation Efficiency||105 - 106 cfu/µg plasmid DNA.|
Q1: What is in this kit and how does it work?
Q2: Does this kit work on C. albicans, S. pombe, or Pichia pastoris?
Q3: How long can I store my competent yeast cells below -70°C?
Q4: How should I thaw the stored frozen competent cells?
Q5: Do the frequencies of freezing and thawing affect the transformation efficiency?
Q6: In step 2 of the transformation, do I need to incubate strictly at 30°C?
Q7: Can I use DNA directly from restriction enzyme digestions without purification?
Q8: How much transformation mixture should I spread on a selective plate?
Q9: Do I need to use plates with sorbitol?
Q10: Do I need to add carrier DNA?
Plasmids were transformed with high efficiency using the Frozen-EZ Yeast Transformation II Kit in the construction of synthetic yeast promoters. Researchers showed that decreasing nucleosome affinity correlated with increasing promoter strength and expression levels higher than native species.Curran KA, et. al. (2014) Design of synthetic yeast promoters via tuning of nucleosome architecture. Nat Commun. 5:4002.
The Frozen-EZ Yeast Transformation II Kit was used to integrate various TEF1 promoter mutants upstream of a gene in Saccharomyces cerevisiae. Efficient transformation allowed comparison of gene activity and, therefore, determination of promoter strength.Nevoigt, E., et al. (2006) Engineering of promoter replacement cassettes for fine-tuning of gene expression in Saccharomyces cerevisiae. Appl Environ Microb, 72(8): 5266-5273.