Qiagen: This is ridiculous

Steven Cohen sbc4 at PO.CWRU.EDU
Fri Aug 30 16:00:52 EST 1996


At 10:37 AM 8/30/96 +0100, you wrote:
>
>> I have never had a problem, one question, what is your bacterial strain? 
>> Also do not overload the column.
>
>Answers:
>strain is DH5-alpha, overnight growth in LB
>As to overloading the column... actually, we found that we IMPROVED the 
>yield from the column if we went above the suggested amount of culture. 
>The protocol says 3ml, but we found we got more plasmid if we used 9 or 10ml.
>
>
If you have got away with loading 9-10 mls then you should consider yourself
lucky. Qiagen yield quickly drop to 0000 if the columns are indeed
significantly overloaded. I have been personally seen this in our lab on
more than one occasion. We also use DH4-a, but in T-broth, giving
significantly more bugs, causing overloading to be a more critical issue.

Steve




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