Yeast Transformation

There are several methods available for transforming yeast cells. For high efficiency transformation I use the Geitz method and add DMSO. Otherwise I just grow some cells and use lithium acetate as described below.
Library Transformations     Scale provides cells for 8 transformations:

add:	
	240 ul 50% PEG 3000
	 36 ul 1 M LiOAc
	 25 ul 2 mg/ml ssDNA
	 50 ul (1 ug or less) plasmid in water
Should Yield 10exp5/ug


Regular Transformation: ( "just to get transformants" but still good efficiency )

	inoculate 5 ml O/N in glass tube
	shake approx 24 hours, 30 C
	take OD 600 (ods are typically 0.5 to 4.0 depending on media)
	take 1 - 1.4 ml cells to eppendorf tube
	pellet cells (spin 4k, 2 min)
	remove sup.
	add:
		240 ul 50% PEG4000
		 36 ul 1M LiOAc, pH 7.5
		 25 ul 2 mg/ml ssDNA
		approx 1 ug plasmid

	vortex, or break up cell pellet by pipetting
	incubate 30 C, 30 minutes
	add 33 ul DMSO
	heat shock 42C, 15 min.
	add 800 ul H20
	vortex
	spin 4k, 2 min.
	resuspend in 100 ul

usually yields 10exp4/ug so plate accordingly


Chris Seidel