AccuPower® HotStart PCR PreMix (with UDG) is applied with Bioneer's enzyme-mediated HotStart patent technology for increasing response specificity and PCR amplification and uracil DNA glycosylase system for eliminating carry-over contamination. Components necessary for PCR such as DNA polymerase, dNTPs, reaction buffer, etc. are thoroughly mixed having vacuum-dried and each packaged with a sufficient amount for one PCR run. In addition, stabilizers are added to maintain enzymatic activities for a month when stored at room temperature (25°C), and for two years when stored in the freezer.
※PreMix product is shipped at room temperature.
※The reaction mixture contains stabilizers that keep full activity of enzymes even at the ambient temperature.
Minimized false positives caused by carryover contamination through the application of Uracil DNA glycosylase system
Excellent sensitivity and amplification efficiency of PCR reactions even with small amounts of DNA
Shortened selection time of PCR condition using PyroHotstart technology, with anyone capable of obtaining of complicated template DNA accurately
Enhanced stability compared to solution-type products allowing delivery at room temperature by adding of stabilizing agents
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
(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.
• gDNA template PCR
• Low-copy target PCR
• Multiple primer pairs PCR
• cDNA template PCR
• Molecular diagnosis
Figure. Comparison of specificity between AccuPower® HotStart PCR PreMix and HotStart PCR PreMix (with UDG).
Specificity test was operated using 7 pairs of primer targeting P53 gene. Reaction mixture was incubated at 37°C for 2 min followed by
95°C for 5 min, 30 cycles of 20 sec at 95°C, 40 sec at 55°C, 1 min at 72°C. AccuPower® HotStart PCR PreMix (with UDG) contains dUTP besides dATP, dGTP, dCTP and dTTP.
The amount of DNA (human) used to test is 10 ng and size of PCR amplicon size each lane is represented below.
Lane 1: 139 bp
Lane 2: 211 bp
Lane 3: 447 bp
Lane 4: 618 bp
Lane 5: 1082 bp
Lane 6: 1296 bp
Lane 7: 1561 bp
M: 100 bp plus DNA ladder (Bioneer Cat. No: D-1030-1)
