|"The important factor considered while applying glass bead method for fungal transformation was the size of the glass beads. The glass beads with 0.45-0.52 mm diameter were found to be suitable for transformation. The ratio of number of spores to the amount of glass beads is also crucial, and the ratio of 1:1 yielded better transformation efficiency. Prior to transformation, the plasmid DNA was linearized with EcoRI restriction enzyme. Polyethylene glycol (PEG 3500) was added to the transformation mixture at a final concentration of 5% wt/ vol and 0.1 M lithium chloride was also added to facilitate the DNA uptake. The use of PEG results into clumping of treated cells which facilitate the trapping of DNA , whereas the use of high concentration of lithium ions renders the cell walls permeable to DNA without the need for protoplasts, a method initially applied for transforming yeast, S. cerevisiae.
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