iTaq™ supermix with ROX gives superior results on the ABI PRISM 7000 sequence detection system. iTaq supermix with ROX (upper) or an alternate universal master mix (lower) was used to amplify tenfold serial dilutions of the 18S rRNA gene, from 100 ng to 1 fg of cDNA from HeLa RNA, using a predesigned assay reagent (PDAR) VIC/MGB probe (Applied Biosystems). Reactions were performed in quadruplicate. The Bio-Rad supermix consistently generated earlier threshold cycle (C_T) values than the alternate mix, as well as better efficiency, even when an ABI-optimized PDAR probe was used for detection.

Comparison of iTaq supermix with ROX and an alternate universal mix.

iTaq™ supermix with ROX exhibits superior performance over six orders of magnitude on the Stratagene Mx3000P. iTaq supermix with ROX (upper) or an alternate universal master mix (lower) was used to amplify tenfold serial dilutions of 100 ng to 100 fg of cyclophilin cDNA from HeLa RNA. A FAM/MGB PDAR probe (Applied Biosystems) was used for detection. The Bio-Rad supermix consistently generated earlier threshold cycle (C_T) values, even when an ABI-optimized PDAR probe was used for detection.

Comparison of iTaq supermix with ROX and an alternate universal mix.

iTaq™ supermix with ROX produces more uniform results in a duplex reaction on the ABI PRISM 7000 sequence detection system. iTaq supermix with ROX (upper) or an alternate universal master mix (lower) was used to multiplex the tubulin and GAPDH targets from 1 ng of cDNA (47 replicates per mix). A FAM-labeled probe was used for tubulin detection, while an ABI PDAR probe labeled with VIC was used to detect GAPDH. In both dye layers, the Bio-Rad mix generated more uniform results with earlier mean threshold cycle (C_T) values than the alternate mix. All reactions were performed in the same 96-well plate.

Comparison of iTaq supermix with ROX and an alternate universal mix.