competent cells again!!

John Augliera jaugliera at RICS.BWH.HARVARD.EDU
Wed Jun 3 14:31:32 EST 1998

	I recently posted a message asking for a protocol and storage
conditions for making bacterial cells competent.   Someone suggested
the procedure in "Maniatis".  I read the protocol which uses successive
washes of cold CaCl2 and the final step calls for flash-freezing the cells in
cold CaCl2 and placing at -70C.  It makes no mention of adding a
cryoprotectant such as glycerol.  Would anyone know if this is OK!! or
would the addition
10% glycerol adversely affect the competency or viability of the cells.


John Augliera
Brigham and Women's Hospital
Boston,MA 02115

email: jaugliera at

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