Transcription of Transformation Protocol Using Heat Shock
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Transformation Protocol Using Heat Shock
Transformation Protocol Using Heat Shock MFT, 11/21/03 1) Take competent cells from 80oC freezer. a. Use DH5 cells in most cases. b. If want to cut at XbaI or other DAM- enzyme site, use SCS110 cells which are deficient in Dam and Dcm methylases. 2) Turn on water bath to 42 C. 3) Put competent cells in a ml tube (Eppendorf or similar). For transforming a DNA construct, use 50 ul of competent cells. For transforming a ligation, use 100 ul of competent cells. You may need more or less cells, depending how competent they are. 4) Keep tubes on ice. 5) Add 50 ng of circular DNA into cells.
Transformation Protocol Using Heat Shock MFT, 11/21/03 1) Take competent E.coli cells from –80oC freezer. a. Use DH5α cells in most cases. b. If want to cut at XbaI or other DAM- enzyme site, use SCS110 cells which are deficient in Dam and …
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