Phusion U Green Multiplex PCR Master Mix, 100 x 50 μL Reactions - FAQs

View additional product information for Phusion U Multiplex PCR Master Mixes - FAQs (F562L, F564S, F562S, F564L)

7 product FAQs found

How much blood can be added directly to multiplex PCR reaction using Phusion U Multiplex PCR Master Mix?

For direct multiplex PCR without prior DNA purification, add up to 4 µL of whole blood to 50 µL final reaction volume.

Can Phusion U Multiplex PCR Master Mix be stored at +4 degrees C?

Yes, Phusion U Multiplex PCR Master Mix can be stored at +4 degrees C for up to 3 months with no effect on performance.

What is the limit of amplicon size using Phusion U Multiplex PCR Master Mix?

Phusion U Multiplex PCR Master Mix is designed for simultaneous amplification of targets up to 2.5 kb.

How many fragments can be amplified in a single reaction using Phusion U Multiplex PCR Master Mix?

We have demonstrated successful simultaneous amplification of 24 DNA fragments.

What are the Phusion U Multiplex PCR Master Mixes and what is the purpose of the Green format?

Thermo Scientific Phusion U Multiplex PCR Master Mixes (Cat. No. F562S, F564S, F562L, F564L) are ready-to-use, 2X end-point PCR master mixes designed for simultaneous amplification of multiple targets in a single tube. Over 20 primer pairs may be combined into a single reaction for highly specific and efficient multiplexing over a broad range of primer and template concentrations. Phusion U Multiplex PCR Master Mixes contains a proprietary buffer with balanced concentrations of all PCR components eliminating the need for tedious optimization.

The Green format of the Phusion U Multiplex PCR Master Mixes (Cat. No. F564S, F564L) further simplifies the workflow: it includes a density reagent and two electrophoresis tracking dyes for direct loading of PCR products on gels.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can I use Phusion Green and Phire Green PCR products for subsequent applications such as sequencing, ligation, and restriction digestion?

Yes. The dyes do not interfere with downstream applications such as DNA sequencing, ligation, and restriction digestion.

What dyes are present in Phusion Green and Phire Green buffers?

Phusion Green and Phire Green buffers contain two dyes for monitoring electrophoresis progress. The blue dye migrates with 3-5 kb DNA fragments and the yellow dye migrates faster than 10 bp DNA fragments in 1% agarose gel. The dyes have excitation peaks at 424 nm and 615 nm, respectively.