Question about maxi plasmid preparation - (Jan/04/2008 )
I prepared plasmid from DH5alpha using the classic alkaloid lysis method (non-kit);
After adding solution I/II/III,centrifugation, 0.6 volume of iso-propyl alcohol,precipitation,washing using 70% alcohol, the protocol said "dissove the precipitates using 3 ml TE buffer and add the same volume of LiCl...."
actually, the TE buffer can not dissove the precipitates completely..
Any one can tell me why?
-glcui-
QUOTE (glcui @ Jan 5 2008, 12:32 AM)
I prepared plasmid from DH5alpha using the classic alkaloid lysis method (non-kit);
After adding solution I/II/III,centrifugation, 0.6 volume of iso-propyl alcohol,precipitation,washing using 70% alcohol, the protocol said "dissove the precipitates using 3 ml TE buffer and add the same volume of LiCl...."
actually, the TE buffer can not dissove the precipitates completely..
Any one can tell me why?
After adding solution I/II/III,centrifugation, 0.6 volume of iso-propyl alcohol,precipitation,washing using 70% alcohol, the protocol said "dissove the precipitates using 3 ml TE buffer and add the same volume of LiCl...."
actually, the TE buffer can not dissove the precipitates completely..
Any one can tell me why?
If the DNA is overdried, then precipitated DNA may not completely dissolve.
-scolix-