The "premix in the kit contains 2.5X concentrations of Tris-HCl, pH 9.0, and MgCl2. The concentration of buffer in the final reaction volume should be 80mM Tris, 2mM MgCl2," according the insert.
I use 4uL of Big Dye, 2uL of the 5X buffer, (and 2uL H20 - since I add 4uL of primer, and 8uL of plasmid DNA for a total of 20uL.) I have also tried the four-fold dilution, which worked for the Pgem standard, but I prefer the half reaction because of the wide variety of samples I receive. tm
>>> "Jan Ilavsky (home account)" <jilavsky@aps.anl.gov> 10/4/99 >>>
Dear colleagues,
I am interested in finding out where I could obtain buffer (called Half Term
Buffer?) used to dilute ABI BigDye kits. Is there a commercial source of
this buffer (in US, or better in Germany)? What is the composition of this
buffer? Can be made in the lab?
Another question is regated to ET-terminator kits from Amersham.
Supposingly, these can also be diluted 4-fold. Is there any experience with
this?
Thank you
Best regards Jan Ilavsky
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Jan Ilavsky
University of Maryland at College Park
Dept. of Chemical Engineering
College Park, MD 20742
USA
Temporary @ Argonne National Laboratory, Advanced Photon Source, Chicago, IL
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