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)
Technical / Specifications
▶ Specifications
|
Enzyme |
Top polymerase |
|
5' to 3' exonuclease activity |
No |
|
3' to 5' exonuclease activity |
No |
|
3'- A overhang |
Yes |
|
Fragment size |
Up to 12 kb |
▶ Experimental data
Figure 1. High sensitivity of AccuPower® Hotstart PCR PreMix
M; 100 bp plus DNA ladder (Bioneer Cat. No: D-1035)
Lane 1; Lamda DNA 10 ng
Lane 2; Lamda DNA 1 ng
Lane 3; Lamda DNA 100 pg
Lane 4; Lamda DNA 10 pg
Lane 5; Lamda DNA 1 pg
Lane 6; Lamda DNA 100 fg
Lane 7; Lamda DNA 10 fg
Lane 8; Lamda DNA 1 fg
Lane 9; Human genomic DNA 10 ng
Lane 10; Human genomic DNA 1 ng
Lane 11; Human genomic DNA 100 pg
Lane 12; Human genomic DNA 10 pg
Figure 2. Comparison of PCR amplification sensitivity between AccuPower® Hotstart
PCR PreMix from Bioneer and other suppliers‘ Hotstart PCR kit.
A) Lamda DNA sensitivity
Lane 1; Lamda DNA 10 ng
Lane 2; Lamda DNA 1 ng
Lane 3; Lamda DNA 100 pg
Lane 4; Lamda DNA 10 pg
Lane 5; Lamda DNA 1 pg
Lane 6; Lamda DNA 100 fg
Lane 7; Lamda DNA 10 fg
Lane 8; Lamda DNA 1 fg
B) Human genomic DNA sensitivity
M; 100 bp plus DNA ladder (Bioneer Cat. No: D-1035)
Lane 1; Human genomic DNA 10 ng
Lane 2; Human genomic DNA 1 ng
Lane 3; Human genomic DNA 100 pg
Lane 4; Human genomic DNA 10 pg
Figure 3. Comparison of PCR amplification sensitivity between AccuPower® Hotstart
PCR PreMix from Bioneer and other suppliers‘ Hotstart PCR kit
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)
Ordering Information
| Cat. No. | Description | Option | Price (AUD) |
|---|---|---|---|
| K-5050 | AccuPower ® HotStart PCR PreMix | 0.2 ml thin-wall 8-strip tubes with attached cap / 96 tubes | $87.69 |
| K-5052 | AccuPower ® HotStart PCR PreMix | 0.2 ml thin-wall 8-strip tubes with attached cap / 96 tubes | $132.31 |
| K-5051 | AccuPower ® HotStart PCR PreMix | 0.2 ml thin-wall 8-strip tubes with attached cap / 480 tubes | $330.77 |
| K-5057 | AccuPower ® HotStart PCR PreMix | 0.2 ml thin-wall 8-strip tubes with attached cap / 480 tubes | $492.31 |
| K-5053 | AccuPower ® HotStart PCR PreMix | 0.5 ml thin-wall tubes with attached cap / 100 tubes | $153.85 |