Sal I cloning problems

Jonas Lindeberg Jonas.Lindeberg at bmc.uu.se
Thu Oct 27 05:40:52 EST 1994


>In article <199410242315.QAA25218 at netcomsv.netcom.com>, ltaylor at genpharm.com
>("Lisa Taylor") writes:
>
>>   OFFICE MEMO          Sal I cloning problems                 Date:10/24/94
>> I have had a great deal of trouble cloning anything which involves a Sal I
>>site.
>>  I would like to know if anyone else has had trouble with this endonuclease
>>and
>> more importantly, if anyone has found a way to digest with Sal I and then get
>> successful ligations.  I have been using Sal I from NEB.
>
>
>I routinely use BSA to 0.15 mg/ml and spermidine to 2 mM in all my restriction
>digests.  One of these (can't remember which) is _essential_ to SalI digests.
>Also, SalI doesn't like being near the end of a DNA fragment - check the NEB
>catalogue reference section for more info.
>
>Richard


????? I cleave with SalI (from boeringer), without any BSA or sperimidine.
It works perfectly well. About the problems with near-end cleavage I agree.

Regards

Jonas





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