YEAST TRANSFORMATION (EPISOMAL)
Use this to transform plasmids into yeast where you do not intend to recombine the DNA into the yeast genome.
0. Before you begin: you need overnights of the yeast strains you will be transforming, use 3ml in YEPD or appropriate media, as well as prepared 10XTE, 50% PEG 3550, salmon sperm DNA, and 1M LiOAC.
0.5: Be sure that you have a 95C and a 42C heat block or water bath powered on and available. People often turn these off at any random time throughout the day, to “save power”, so doublecheck! YOU CANNOT WARM THE BLOCK OR BATH WITH YOUR CELLS IN IT IT MUST BE PRE-WARMED. It's a HEAT SHOCK
|Number of Reactions||1||5||10||20|
|ul to use of:|
|10X TE Buffer||20||100||200||400|
|50% PEG 3350**||160||800||1600||3200|
|Salmon Sperm DNA, freshly boiled*||5||25||50||100|
- Boiled means means put the salmon sperm DNA in a 95C heat block before use. The block is on Katie's bench and the SSDNA is in -20C #2 in a labeled box- take an aliquot and discard ay unused after boiling
- PEG 3550 is Sigma P/N 3640