About QuestTaq™ PreMix
QuestTaq™ PreMix is supplied as a convenient 2X concentrated “master mix” containing all the reagents (i.e., dNTPs, MgCl2, and enhancers) necessary for robust PCR with little or no by-product formation. The QuestTaq™ PreMix has been optimized for the non-biased amplification of cystosine, 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), and glucosyl-5-hydroxymethylctosine (g5hmC) containing DNA, ensuring high yield amplification across a wide range of templates. The QuestTaq™ PreMix differs from QuestTaq™ qPCR PreMix (Cat. Nos. E2052 & E2053 from Zymo Research) in that it excludes SYTO® 9 dye from the PreMix solution, making it compatible with real-time and quantitative PCR with fluorescent dyes of the researcher’s choosing.
|Storage||QuestTaq™ PreMix should be stored between -20 °C to -80 °C for maximum performance. Performance is guaranteed for 1 year from the time of receipt.|
|Unit Definition||One unit (U) enzyme of QuestTaq™ DNA polymerase is defined as the amount of enzyme required for the incorporation of 10 nmol dNTPs into an acid-insoluble form in 30 minutes at 72°C.|
|Enzyme Concentration||Reaction conditions at 1X (20 µl total volume) will contain 2 units of QuestTaq™ DNA polymerase.|
QuestTaq™ polymerase consistently yields robust amplicons from DNA templates having modified/unmodified cytosines. The figure shows the level (intensity) of an ~900 bp product generated from DNA templates using QuestTaq™ PreMix or the polymerases from Suppliers X and Y. In each case, PCR products were separated in a 0.8% (w/v) agarose/TAE/EtBr gel. Lanes correspond to amplicons from template DNA containing: unmodified cytosine (c), 5-methylcytosine (m), 5-hydroxymethylcytosine (h), or glucosyl-5-hydroxymethylcytosine (g). (M) is a 1 kb DNA Marker (Zymo Research, Cat. No. M5006-50).