Application

Cloning into vectors, library construction, TA cloning, linker ligation, and re-circlization of linear DNA

Figure 1. Stability test of AccuPower DNA Ligation PreMix at room temperature.

 

Lane 1 – 12: Lambda DNA / Hind lll fragment (1 µg)
Lane 12 – 24: Lambda DNA / EcoR V fragment (1 µg)
Lane 2, 3, 14, 15: Ligation with AccuPower Ligation PreMix stored at room temperature for 1 month
Lane 5, 6, 17, 18: Ligation with AccuPower Ligation PreMix stored at room temperature for 2 months
Lane 8, 9, 20, 21: Ligation with AccuPower Ligation PreMix stored at room temperature for 3 months
Lane 11, 12, 23, 24: Ligation with AccuPower Ligation PreMix stored at room temperature for 4 months

Figure 2. DNA Ligation efficiency comparison between AccuPower DNA Ligation PreMix and other competitors’ products.

 

Lane 1, 9: Intact Lambda DNA (1 µg)
Lane 2 – 8: Lambda DNA / Hind lll fragment (1 µg)
Lane 10 – 16: Lambda DNA / EcoR V fragment (1 µg)
Lane 3, 4, 11, 12: Ligation with AccuPower Ligation PreMix
Lane 5, 13: Ligation with T4 DNA Ligase from company N
Lane 6, 14: Ligation with Quick Ligation Kit from company N
Lane 7, 15: Ligation with LigFast Rapid DNA Ligation System from company P
Lane 8, 16: Ligation with Ready-To-Go T4 DNA Ligase from company A

Specifications

5' to 3' exonuclease activity: Yes
3' to 5' exonuclease activity: No
Terminal transferase activity: Yes
Fragment Size: 1 kb

Application

- Gene expression profiling
- Target DNA quantification
- Microbial detection
- SNP analysis
- Viral/bacterial pathogen load determination
- Evaluation of primer pair performance for probe-based real-time PCR

Figure 1. Real-Time PCR Data of AccuPower® DualStar™ qPCR PreMix
AccuPower® DualStar™ qPCR PreMix provides at least 7 orders of magnitude in dynamic range (10⁷ ~ 10¹copies/reaction).
A: Amplification curve. West Nile Virus (WNV) primers and TaqMan-based probe were added into DualStar qPCR Premix. A series of WNV positive control diluents were tested.
B: Standard curve. All data were obtained using Exicycler™ 96 Real-Time Quantitative PCR System (Gentaur).

Figure 2. Data using Various kinds of Real-time PCR Instruments
AccuPower® DualStar™ qPCR PreMix provides at least 7 orders of magnitude in dynamic range (10⁷ ~ 10¹copies/reaction).
A: Amplification curve. and standard curve using ABI 7500 Format Fast Real-time PCR machine (Applied Biosystems). A series of WNV positive control diluents were tested.
B: Amplification curve and standard curve using ABI 7500 Format Real-time PCR machine (Applied Biosystems). 
C: Amplification curve and standard curve using Opticon Real-time PCR machine (MJ Research, currently Bio-Rad).

Figure 3. Comparison of detection sensitivity between AccuPower® DualStar™ qPCR PreMix and other supplier's master mixture
West Nile Virus (WNV) primers and TaqMan-based probe were added into AccuPower® DualStar™ qPCR Premix and other supplier's master mixture. A series of WNV positive control diluents were tested. Reaction mixtures were prepared and qPCR was performed according to each supplier's protocol. All data were obtained using ABI 7500 Format Fast Real-time PCR machine (Applied Biosystems).

Specifications

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment size: 6 kb

Application

cDNA synthesis followed by PCR

Figure 1. Specific amplification of 5' - UTR region of HCV with AccuPower RT-PCR PreMix. Human DNA was amplified to generate 0.1-1.5 kb PCR fragments using 1 U of polymerase. The comparison was performed under the same experimental conditions.

 

Lane 1: 123 DNA Ladder
Lane 2: Negative control
Lane 3, 4: HCV positive serum
Lane 5: HCV negative serum

Specifications

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment size: 9 kb

Application

cDNA synthesis

Figure 1. Reliability and reproducibility test with AccuPower RT PreMix. Each template showcases amplified target genes.

 

Lane M: 100 bp DNA Ladder (D-1030)
Lane 1-4: Reliability test of each lot with AccuPower RT PreMix

Specifications

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: Yes
Fragment Size: Up to 1 kb

Application

Target	Application
Human and Animal
	STR analysis for determining genetic profiles in forensic cases
	Molecular diagnostic analysis
	Genotyping assay
	Qualitative and semi-qualitative gene expression assay
	Mutant screening
	Transgenic organism analysis
Plant
	STR analysis
	Detection of pathogens/bacterial infection
	Transgenic organism analysis
	Qualitative and semi-qualitative gene expression assay

 

Figure 1. Single PCR and Multiplex PCR using AccuPower Gold Multiplex PCR PreMix
Each lane from left to right indicates the single and multiplex PCR products using AccuPower Gold Multiplex PCR PreMix. The last lane labeled Multi represents all amplicons amplified in a single reaction. 
a) 10-plex PCR, b) 20-plex PCR

 

Figure 2. High specificity of AccuPower Gold Multiplex PCR PreMix
Each lane from left to right represensts the progressive number of primer sets (1 – 20) included in single AccuPowerGold Multiplex PCR PreMix reactions.

 

Figure 3. Comparison of amplification quality between AccuPower Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.
6-plex (a), 10-plex (b), 20-plex (c) primers were added into AccuPower Gold Multiplex PCR PreMix and competitor master mixtures. A series of human genomic DNA diluents were tested (Lane 1: 100 ng, Lane 2: 10 ng, Lane 3: 1 ng).

 

Figure 3. Comparison of amplification quality between AccuPower Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.
6-plex (a), 10-plex (b), 20-plex (c) primers were added into AccuPower Gold Multiplex PCR PreMix and competitor master mixtures. A series of human genomic DNA diluents were tested (Lane 1: 100 ng, Lane 2: 10 ng, Lane 3: 1 ng).

 

Figure 4. Comparison of amplification quality, using a Caliper Labchip® 90 system, between AccuPower Gold Multiplex PCR PreMix and competitor Multiplex PCR kits.
a) Virtual Gel Image. Gel image illustrates data reproducibility of the Caliper LabChip® 90 system.
b) Overlay of expression levels using Gentaur’s AccuPower Gold Multiplex PCR PreMix and competitor multiplex PCR kits. The electropherogram displays the data between 10 PCR product yields using a 10-plex primer set to illustrate amplification efficiency.
c) The graph shows the total concentration of PCR products between AccuPower Gold Multiplex PCR PreMix and other supplier’s master mixtures.

Specifications

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: Yes
3’ – A Overhang: No
Fragment Size: 20 kb

Application

Gene synthesis, PCR or primer extension requiring high fidelity, blunt end PCR cloning or mutagenesis, any amplification requiring high fidelity

Figure 1. Comparison of long PCR amplification between AccuPower TLA PCR PreMix and 2 high-fidelity PCR enzymes. Reaction mixtures were prepared and PCR cycling conditions were performed following the protocol recommendations.

 

Lane M1: 1 kb DNA Ladder (D-1040)
Lane M2: Lambda/Hind lll Marker (D-1050)
Lane 1, 8, 15: 2 kb PCR product
Lane 2, 9, 16: 5 kb PCR product
Lane 3, 10, 17: 8 kb PCR product
Lane 4, 11, 18: 10 kb PCR product
Lane 5, 12, 19: 12 kb PCR product
Lane 6, 13, 20: 15 kb PCR product
Lane 7, 14, 21: 20 kb PCR product

Figure 2. Comparison of amplification efficiency of AccuPower TLA PCR PreMix and other equivalent products PCR was carried out using AccuPower TLA PCR PreMix and 3 high-fidelity PCR enzymes from other companies. Parallel reactions were performed following the protocol recommendations, using 100 ng, 10 ng, 1 ng and 100 pg human genomic DNA. A 1 kb fragment of the human IR gene was amplified in 30 PCR cycles.

 

M: 100 bp DNA Ladder (D-1030)
Lane 1, 6, 11, 16: 100 ng of human genomic DNA
Lane 2, 7, 12, 17: 10 ng of human genomic DNA
Lane 3, 8, 13, 18: 1 ng of human genomic DNA
Lane 4, 9, 14, 19: 100 pg of human genomic DNA
Lane 5, 10, 15, 10: Negative control template

Figure 3. . Comparison of detection sensitivity between AccuPower TLA PCR PreMix, Taq DNA polymerase and 2 high-fidelity PCR enzymes in amplification of various sizes. Reaction mixtures were prepared and PCR cycling conditions were performed following the protocol recommendations.

 

M: 1 kb DNA Ladder (D-1040)
Lane 1, 6, 11, 16: 2 kb PCR product
Lane 2, 7, 12, 17: 4 kb PCR product
Lane 3, 8, 13, 18: 5 kb PCR product
Lane 4, 9, 14, 19: 7 kb PCR product
Lane 5, 10, 15, 10: 8 kb PCR product

Figure 4. . A 2 kb Lambda DNA fragment was amplified according to the protocol recommendations using various extension times ranging from 20 sec to 2 min.

 

M: 1 kb DNA Ladder (D-1040))
Lane 1, 6, 11, 16: 20 seconds of extension time
Lane 2, 7, 12, 17: 40 seconds of extension time
Lane 3, 8, 13, 18: 1 minute of extension time
Lane 4, 9, 14, 19: 1 minute 30 seconds of extension time
Lane 5, 10, 15, 10: 2 minutes of extension time

Specifications

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
Terminal transferase activity: Yes
Fragment Size: Up to 12 kb

Application

HotStart PCR up to 12 kb
PCR with complex genomic templates/low copy
Templates cDNA
Multiplex PCR reactions

Figure 1. Specificity comparison between AccuPower PCR PreMix and AccuPower HotStart PCR PreMix. 50 ng human genomic DNA was amplified into 2.5 - 3 kb fragments with the two products.

 

A: AccuPower PCR PreMix
B: AccuPower HotStart PCR PreMix
Lane M: 1 kb DNA Ladder (D-1040)
Lane 1: 2.5 kb product
Lane 2: 3.0 kb product

Figure 2. Specificity comparison of multiplex PCR with various HotStart PCR reagents. Each PCR mixture with 2 ng of human genomic DNA and 2 pairs of primers (p53 + p55 and p55 + p63) was incubated for 2 hours at 37°ree;C prior to performing multiplex PCR.

 

M: 100 bp DNA Ladder (D-1030)
A: AccuPower HotStart PCR PreMix
B: Control – AccuPower PCR PreMix
C: Company T's antibody mediated HotStart Taq DNA polymerase (0.5 U)
D: Company A’s antibody mediated HotStart Taq DNA polymerase (1.0 U)

Specifications

5' to 3' exonuclease activity: Yes
3' to 5' exonuclease activity: No
3' – A Overhang: Yes
Fragment size: Up to 5 kb from human genomic DNA and 10 kb from Lambda DNA

Application

- High specificity PCR
- High sensitivity PCR
- Low-copy target PCR 
- Multiplex PCR
- cDNA amplification
- TA cloning

Figure 1. Comparison of PCR amplification specificity between AccuPower® PyroHotStart Taq PCR PreMix from Gentaur and other suppliers' Hot start PCR master mix.

PCR reactions were performed according to each supplier's protocol. The PrP gene was amplified from human genomic DNA with two different primer sets, separately. This data shows that AccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency and specificity than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder(Gentaur, Cat. No.D-1030)
Lane 1: 100 ng DNA, PrP primer set (500 bp)
Lane 2: 10 ng DNA, PrP primer set (500 bp) 
Lane 3: 100 ng DNA, PrP primer set (705 bp)
Lane 4: 10 ng DNA, PrP primer set (705 bp)

Figure 2. Comparison of PCR amplification specificity between AccuPower PyroHotStart Taq PCR PreMix from Gentaur and other suppliers' Hot start PCR master mix.

The ApoE gene was amplifed from 100 ng of human genomic DNA (The PCR product size is 268bp). This data shows thatAccuPower PyroHotStart Taq PCR PreMix has higher amplificition efficency and specificity than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Gentaur, Cat. No. D-1030)
Lane 1: AccuPower PyroHotStart Taq PCR PreMix
Lane 2: Supplier I Hotstart Taq PCR preMix
Lane 3: Supplier S Hotstart Taq PCR masterMix
Lane 4: Supplier T Hotstart Taq PCR masterMix
Lane 5: Supplier Q Hotstart Taq PCR masterMix

Figure 3. AccuPower PyroHotStart Taq PCR PreMix has high amplification efficiency and specificity.

Specificity test was performed using 7 different sets of primers targeting the P53 gene. 10 ng of human genomic DNA was used for each PCR reaction. The cycling conditions were 95°C for 5min, 30 cycles of 95°C for 20 sec, 55°C for 40 sec, and 72°C for 1 min, and 72°C for 5 min for final extension.
Lane M: 100bp DNA Ladder (Gentaur, Cat. No. D-1030)
Lane 1: P75/73 primer set (139 bp)
Lane 2: P55/53 primer set (211 bp)
Lane 3: P55/63 primer set (447 bp)
Lane 4: P75/83 primer set (618 bp)
Lane 5: P55/73 primer set (1082 bp)
Lane 6: P65/83 primer set (1296 bp)
Lane 7: P55/83 primer set (1561 bp)

Figure 4. Comparison of PCR amplification between AccuPower PyroHotStart Taq PCR PreMix from Gentaur and other suppliers' Hot start PCR master mix.

Sensitivity test was performed by amplifying the IRGC gene from a serial dilution of human genomic DNA. This data shows that AccuPower PyroHotStart Taq PCR PreMix has higher amplification efficiency than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Gentaur, Cat. No. D-1030)
Lane 1: 10 ng human genomic DNA
Lane 2: 1 ng human genomic DNA
Lane 3: 100 pg human genomic DNA
Lane 4: 10 pg human genomic DNA

Figure 5. Comparison of cDNA template amplification between AccuPower PyroHotStart Taq PCR PreMix and other suppliers' Hot start PCR master mix.

cDNA synthesized from 10-fold serial-diluted human total RNA from 10 ng to 10 pg using AccuPower RocketScript™ Cycle RT PreMix (Gentaur, Cat. No. K-2201) was used as a template for PCR amplification. This data shows thatAccuPower® PyroHotStart Taq PCR PreMix has higher amplification efficiency than other suppliers' Hot start PCR master mix.
Lane M: 100 bp DNA Ladder (Gentaur, Cat. No. D-1030)
Lane 1: 10 ng human total cDNA
Lane 2: 1 ng human total cDNA
Lane 3: 100 pg human total cDNA
Lane 4: 10 pg human total cDNA

 

Specifications

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
Terminal transferase activity: Yes
Fragment Size: Up to 10 kb

Application

Standard PCR

Figure 1. Processivity comparison with Lambda DNA template and different PCR reagents.

 

M: 100 bp Plus DNA Ladder (D-1035)
A: Accupower PCR PreMix
B: Taq mastermix from Company S
C: Taq mastermix from Company T
Lane M: 1 kb DNA Ladder (D-1040)
Lane 1: 1 kb PCR product
Lane 2: 2 kb PCR product
Lane 3: 3 kb PCR product
Lane 4: 4 kb PCR product

Figure 2. Comparison of sensitivity test for AccuPower PreMix PCR master mix and other companies' products using serial diluted human genomic DNA.

 

Reaction condition: 95°C for 5 minutes, followed by 35 cycles of 20 seconds at 95°C, 20 seconds at 60°C, 30 seconds at 72°C.

A: human Insulin receptor gene
B: DNA cross-link repair 1A gene
I: AccuPower PCR PreMix
II: Taq DNA Polymerase from Company A
III: Taq DNA Polymerase from Company Q
IV: PCR PreMix from Company I
M: 100 bp DNA ladder (D-1030)
Lane 1: 10 ng
Lane 2: 1 ng
Lane 3: 100 ng
Lane 4: 10 ng