Platinum™ Direct PCR Universal Master Mix, 100 reactions - FAQs

View additional product information for Platinum™ Direct PCR Universal Master Mix - FAQs (A44647500, A44647100, A44647200)

9 product FAQs found

Can I use plant or tissue sample that is not on the list of the Platinum Direct PCR Universal Master Mix tested samples?

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.

When performing Direct PCR with the Platinum Direct PCR Universal Master Mix, do I need to use Proteinase K?

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.

Is the PCR product from reactions performed with Platinum Direct PCR Universal Master Mix compatible with E-Gel agarose gels?

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.

With the Platinum Direct PCR Universal Master Mix, can I leave the samples in the Lysis Solution (Lysis Buffer containing Proteinase K) for longer than 1 min?

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.

What is the biggest sample size that I can use with Platinum Direct PCR Universal Master Mix?

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.

With the Platinum Direct PCR Universal Master Mix, can I adjust the volume of Lysis Buffer depending upon the sample size?

If your sample is larger than 1 mm in diameter, we recommend increasing the Lysis Buffer volume to make sure that the sample is completely covered in the buffer. Add Proteinase K accordingly.

Does the Platinum Direct PCR Universal Master Mix come with a tissue puncher? What kind of tools do I need for sample handling?

No, a tissue puncher is not provided with the kit. If a tissue puncher is not available, cut the sample with a scalpel. Tweezers, scalpel and other tools need to be cleaned with 2% NaClO solution to avoid cross contamination.

What are your recommendations for obtaining a sample from plant or animal tissue for amplification with Platinum Direct PCR Universal Master Mix?

In order to obtain best results when amplifying DNA directly from a sample using Platinum Direct PCR Universal Master Mix, it is important to use a very small amount of starting material. The recommended size is 1-2 mm in diameter. Possible tools for sample handling are: tissue puncher, scalpel or 100 µL pipette tip for soft samples.

Which FFPE sample types can I use with Platinum Direct PCR Universal Master Mix for direct amplification from tissue samples?

FFPE tissue, FFPE tissue sections, FFPE tissue blocks and microscope slides can be used with Platinum Direct PCR Universal Master Mix. DNA is highly fragmented in FFPE samples, and DNA quality being one of the main factors for successful amplification, we recommend amplifying fragments up to 300 bp with this master mix.