Platinum™ Direct PCR Universal Master Mix

You can use any plant or tissue sample. However, when working with new sample materials, we recommend using the Lysis protocol as it allows several PCR reactions to be performed from the same sample during optimization. We also recommend having a positive control with purified sample DNA to ensure that the PCR conditions are optimal. If the positive control with purified DNA fails, the PCR conditions should be optimized before continuing further.

Proteinase K is required when PCR is performed directly from tissue samples using the Direct PCR protocol. Cell debris present in these PCR products can cause DNA fragments to get trapped in the agarose gel wells and Proteinase K helps to eliminate this.

Electrophoretic separation on E-Gel agarose gels depends on the salt concentration in the analyzed sample. For optimal band separation, we recommend diluting PCR reactions 2- to 20-fold with water, prior to running on E-Gel agarose gels. The dyes in the Platinum Direct PCR Universal Master Mix do not interfere with fragment separation on E-Gel agarose gels.

Samples can be incubated for up to 8 hr at room temperature in the Lysis Solution, prior to the 98 degrees C Proteinase K inactivation step. Please note that longer incubation may increase the product yield. For longer incubation, the Proteinase K inactivation step may be increased up to 10 min.

For the Direct protocol, the recommended sample size is up to 1 mm. For the Lysis protocol, the recommended sample size is up to 2 mm. Sample size can be increased up to 1 cm, however, Lysis Buffer volume needs to be increased to cover the whole sample.