This product is a key reagent for yeast transformation, consisting of Tris, EDTA, PEG3350, and lithium acetate in deionized water, with the TE-lithium acetate component sterilized by autoclaving. The three components (A:B:C) are used in an 8:1:1 ratio for the preparation of yeast competent cells and exogenous DNA transformation.
Mechanism of Action
• PEG3350 (polyethylene glycol, MW 3,350): In the presence of high concentrations of lithium acetate, PEG protects the yeast cell membrane from excessive damage, while promoting closer contact between plasmid DNA and the cell membrane, significantly enhancing transformation efficiency. Studies have shown that PEG3350 can achieve transformation efficiencies of up to 10⁵ cfu/μg DNA. PEG with a molecular weight of approximately 3,000 exhibits optimal transformation-promoting activity, and PEG3350 was selected based on this principle.
• Lithium Acetate (LiAc): Induces a transient competent state in yeast cells, increasing membrane permeability and enabling the uptake of exogenous DNA.
• TE Buffer (Tris-EDTA): Maintains pH stability, while EDTA chelates metal ions to protect DNA from nuclease degradation.
Important Note
The concentration of PEG is critical for transformation efficiency. Strict adherence to the recommended ratio is essential to ensure consistent and reproducible results.
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