Product No. 12140306001
Template quality significantly influences PCR outcome.
For the amplification of GC-rich targets up to 5 kb in length, DNA with repetitive sequences, and DNA mixtures with varying GC content, Roche recommends using the GC-RICH PCR System or the GC-RICH PCR System, dNTPack. The GC-RICH PCR System includes three specially formulated components not included in most PCR systems:
(1) Special enzyme mix and (2) reaction buffer, both containing detergents and DMSO;(3) Special GC-RICH Resolution Solution.
The optimal reaction conditions (incubation times and temperatures, concentration of GC-RICH PCR System enzyme mix, GC-RICH resolution solution, template DNA, Mg2+) depend on the template/primer pair and must be determined for each assay. For difficult templates the titration of GC-RICH Resolution Solution may be required (0.5 to 2.5 M, in steps of 0.25 M).
In addition to the above, also consider the following options to optimize PCR using difficult templates: