An Economic PCR Enhancer for GC-Rich PCR Templates
|Author: Ralser, et al.|
|Affiliation: Max Planck Institute for Molecular Genetics|
|Source: Protocol Online|
|Date Added: Mon Feb 02 2009|
|Date Modified: Sat Mar 14 2009|
|Abstract: An combinatorial enhancer solution (CES) which efficiently improves many PCR using different polymerases.|
Since PCR is often problematic on GC-rich templates, we have evaluated different PCR enhancing additives and generated an Combinatorial Enhancer Solution (CES). We tested this solution on approx. 50 different primer pairs using several different polymerases
The 5x concentrated Combinatorial Enhancer Solution (CES) contains:
2.7 M betaine
6.7 mM DTT
6.7% DMSO and
55 μg/ml BSA
Add this solution to your PCR and see if it works
Some hints if your PCR still doesn't work.
We tested this enhancer also on PCR which works fine without additive and we virtually observed no negative effects. Therefore, we are now adding this enhancer for all our routine PCR.
65 mM Tris–HCl
16.6 mM (NH4)2SO4,
3.1 mM MgCl2
0.01% (v/v) Tween 20
pH 8.8 (or 8.0)
and supplemented with 5-10 pico-moles of each oligo (per reaction) and 0.5 μl of 25mM dNTP mix.
If this enhancer helps you to improve your experiments and helps you to save time and money, be fair and cite the work:
M. Ralser, R. Querfurth, H.J. Warnatz, H. Lehrach, M.L. Yaspo and S. Krobitsch An efficient and economic enhancer mix for PCR. Biochem Biophys Res Commun. 2006 Sep 1;347(3):747-51. Epub 2006 Jul 5