This mix contains an engineered DNA polymerase in an optimzed reaction buffer and ultrapure dNTPs. HiDi DNA polymerase is hotstart-formulated with an aptamer. Temperatures above 50-55°C cause the aptamer’s secondary structure to melt and will set-free the polymerase.
HiDi DNA polymerase can also be used for real-time cycling, when adding a suitable real-time dye, for example GreenDye (#2000).
However, this mix can not be used for hydrolysis-based probes (e.g. TaqMan) as the 5'-3' nuclease is not active. For this application see HiDi Taq polymerase (#9201).
This product is shipped on cool packs. It is recommended to store the product upon arrival at -20°C.
HiDi 2x PCR Master Mix is tested for successful ASA performance detecting a genomic SNP (rs72921001) in HeLa genomic DNA. PCR products were subsequently analysed on a 2.5% agarose gel. Specific product was visualized by ethidium bromide staining at the correct amplicon length of 109 bp for the matched primer. In case of mismatch primer no product formation was observed after 45 cycles. The activity of HiDi DNA polymerase was monitored and adjusted to a specific DNA polymerase activity using an artificial DNA template and a DNA primer. Enzyme concentration was determined by protein-specific staining. Please inquire more information at [email protected] for the lot-specific concentration. No contamination was detected in standard test reactions.