Endofree Maxiprep and TOP10 cells

Bertrand Collet b.collet at abdn.ac.uk
Thu Jun 29 04:10:45 EST 2000


Hi everybody,

I am having some problem of DNA quality with the Endofree Maxiprep
Plasmid purification kit: A260/A280 ratio are never above 1.4-1.5 when
this protocole is supposed to give ratios above 1.8. I tried to reduce
the volume of the starting culture, use twice reagent for the alkaline
lysis ... there is no way to improve the DNA quality. Are the TOP 10
cells in cause (high carbohydrate content ...) ? Is it worth changing
bacteria strain ? Is there anybody who had this problem with TOP10
strain ?
I do need top quality DNA for transfection purpose.

Thanks for your help,


Dr Bertrand Collet
University of Aberdeen
Department of Zoology
Fish Immunology Group
Tillydrone avenue
Aberdeen AB24 2TZ
Scotland
Phone: (+44) (0) 1224 273 796
Fax: (+44) (0) 1224 272 396







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