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AccuPower® HotStart PCR PreMix
AccuPower® HotStart PCR PreMix enhances response specificity and PCR amplification by applying Bioneer's enzyme-mediated HotStart patent technology. The components necessary for PCR such as DNA polymerases, dNTPs, reaction buffers, etc. are thoroughly mixed and vacuum-dried, each packaged with amounts sufficient for a single PCR run. In addition, stabilizers are added to keep the activity stable for one month when stored at room temperature (25°C) and for two years when stored in a freezer.
※PreMix product is shipped at room temperature.
※The reaction mixture contains stabilizers that keep full activity of enzymes even at the ambient temperature.
Features and Benefits
- Specificity
Effective amplication even with small amounts of template DNA by using Bioneer's differentiated PyroHotstart technology to minimize products from non-specific amplification and maximize efficiency of PCR reaction.
- Sensitivity
Excellent sensitivity and amplification efficiency even with small amounts of DNA
- Stability
Enhanced stability allowing delivery at room temperature compared to solution-type products by addition of stabilizing agents
- User-friendly
Shortened optimization time for PCR conditions and accurate acquisition of complicated template DNA with PyroHotstart technology
- Reproducibility
Minimized deviation between lots and reproducible results even for replicates tested repeatedly under same conditions and variables by a mass production under ISO 9001 quality system
Overview
(a) Inhibition of PCR reaction in the zero cycle
After preparing the PCR mixture, PPi is used to prevent the formation of non-specific reactants before reaching the pre-denaturation temperature.
Also, since DNA polymerases requires Mg ions to activate, PPi, having high affinities with Mg2+ ions, traps them to inhibit the PCR.
(b) Denaturation
PPase included in the mixture is activated when the temperature rises during the denaturation step to decompose PPi into Pi for dissociating the Mg ions.
The isolated ions bind with DNA polymerases to activate, allowing the PCR reaction to proceed.
(c) Annealing & Extension
The activated DNA polymerases amplifies the target DNA as the PCR cycle progresses.
Application
• High specificity PCR
• High sensitivity PCR
• gDNA template PCR
• Low - copy target PCR
• Multiple primer pair PCR
• cDNA template PCR
Experimental data
Figure. Specificity comparison between AccuPower® PCR PreMix and AccuPower® HotStart PCR PreMix
Lane 1: P75/P73 primer set (139 bp)
Lane 2: P55/P53 primer set (211 bp)
Lane 3: P55/P63 primer set (447 bp)
Lane 4: P75/P83 primer set (618 bp)
Lane 5: P55/P73 primer set (1082 bp)
Lane 6: P65/P83 primer set (1296 bp)
Lane 7: P55/P83 primer set (1561 bp)