TaqMan® Genotyping Master Mix can be used in place of TaqMan® Universal PCR Master Mix for end-point fluorescent detection in SNP genotyping applications. This master mix optimizes the preferential binding of the allele-specific probe, providing exceptional separation and clustering of alleles, and consistently strong fluorescent signals. TaqMan® Genotyping Master Mix provides unambiguous allelic discrimination even for the most challenging templates, such as GC-rich regions.
TaqMan® Genotyping Master Mix is supplied at a 2X concentration, which is comprised of AmpliTaq Gold® DNA Polymerase UP (Ultra Pure), dNTPs, optimized buffer components, and a passive internal reference based on proprietary ROX™ dye. It is available in several sizes ranging from 10 mL to 100 mL. Recommended storage conditions are 2-8ºC, so thawing is not required before use.
The stability of TaqMan® Genotyping Master Mix, which utilizes a hot-start enzyme, allows for flexibility and high throughput. It is stable at room temperature pre- and post-PCR, which allows for easy bench top set-up and the ability to carry experiments out over multiple days.
I have been using TaqMan® Universal PCR Master Mix with Applied Biosystems’ TaqMan® SNP Genotyping Assays for several years. I have had good results, but there were always a few assays that required further optimization and/or I could never get to cluster well. When the new TaqMan® Genotyping Master Mix product came out I heard it would provide superior results, particularly for assays with challenging templates. I was pleasantly surprise at just how amazing the results were. I was using the TaqMan® Universal PCR Master Mix to genotype a particular assay and I didn’t get any cluster separation. Attempts at optimization by both me and other collaborative labs did not improve the assay’s performance. I then used the new TaqMan® Genotyping Master Mix on the same assay, using standard protocols, and the results were like night and day. I had discrete clusters and nearly perfect call rates. We have, since that time, experienced the same scenario with multiple assays.