Upon linearization of plasmid DNA for electroporation mediated transformation in yeast, is it necessary to precipitate out the DNA from the restriction reaction via alcohol (With 75% ethanol). Isn't it possible to just deactivate the restriction enzyme via heat treatment and proceed towards transformation using the same restriction mixture?
Heat treatment would also denature your dsDNA into ssDNA. Second the buffer used for the restriction digest might affect your transformation efficiency. Third you only use 70-75% ethanol to wash the pellet. Use pure isopropanol 1.5x v/v to do the precipitation. It also helps to chill your solution for 1-2h to improve precipitation yields.
In addition to what Robert Adolf Brinzer writes, adding some carrier such as tRNA helps to precipitate your DNA if it is at low levels.
Regarding using ethanol, it works but you need a final concentration of around 75%, in other words adding 3 volumes of 100% ethanol to your solution.
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