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 GENTAUR Europe BVBAVoortstraat 49, 1910 Kampenhout BELGIUM Tel 0032 16 58 90 45 Fax 0032 16 50 90 45 This email address is being protected from spambots. You need JavaScript enabled to view it.">This email address is being protected from spambots. You need JavaScript enabled to view it. |
GENTAUR BULGARIA
53 Iskar Str. 1191 Kokalyane, Sofia
Tel 0035924682280
Fax 0035929830072
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GENTAUR France SARL
9, rue Lagrange, 75005 Paris
Tel 01 43 25 01 50
Fax 01 43 25 01 60
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GmbH Marienbongard 20
52062 Aachen Deutschland
Tel (+49) 0241 56 00 99 68
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GENTAUR Ltd.
Howard Frank Turnberry House
1404-1410 High Road
Whetstone London N20 9BH
Tel 020 3393 8531
Fax 020 8445 9411
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GENTAUR Poland Sp. z o.o.
ul. Grunwaldzka 88/A m.2
81-771 Sopot, Poland
Tel 058 710 33 44
Fax 058 710 33 48
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GENTAUR Nederland BV
Kuiper 1
5521 DG Eersel Nederland
Tel 0208-080893
Fax 0497-517897
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GENTAUR SRL IVA IT03841300167
Piazza Giacomo Matteotti, 6, 24122 Bergamo
Tel 02 36 00 65 93
Fax 02 36 00 65 94
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GENTAUR Spain
Tel 0911876558
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Genprice Inc, Logistics
547, Yurok Circle
San Jose, CA 95123
Phone/Fax:
(408) 780-0908
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GENPRICE Inc. invoicing/ accounting:
6017 Snell Ave, Suite 357
San Jose, CA. 96123
Serbia, 
Macedonia,
Montenegro, 
Croatia:
Tel 0035929830070
Fax 0035929830072
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GENTAUR Romania
Tel 0035929830070
Fax 0035929830072
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GENTAUR Greece
Tel 00302111768494
Fax 0032 16 50 90 45
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Other countries
Luxembourg +35220880274
Schweiz Züri +41435006251
Danmark +4569918806
Österreich +43720880899
Ceská republika Praha +420246019719
Ireland Dublin +35316526556
Norge Oslo +4721031366
Finland Helsset +358942419041
Sverige Stockholm +46852503438
Magyarország Budapest +3619980547
AccuPower CycleScript RT PreMix
AccuPower CycleScript RT (available with: dT20, dN12, or dN6 primers) PreMix is an easy to resuspend lyophilized PCR Master Mix of CycleScript Reverse Transcriptase, a primer, and all of the other components for cDNA synthesis conveniently packaged in individual tubes. Simply add template RNA, and water. Then, the reverse transcription is performed by either a Cyclic RT reaction (patent pending) or conventional reverse transcription PCR. The use of Cyclic RT produces cDNA amplification and better results compared to conventional reverse transcription PCR – especially for rare transcripts. The resulting cDNA can be used in a variety of applications such as reverse transcription PCR (RT-PCR). The AccuPower CycleScript RT (dT20, dN12, or dN6) PreMix series is stable for 2 years at - 20°C due to a patented stabilizer.
AccuPower CycleScript RT was developed for both a conventional reverse transcription PCR at a fixed temperature at 42°C as well as Cyclic Reverse Transcription that is carried out like a PCR (please see the following comparison table). Gentaur’s Cyclic RT is an innovative technology to synthesize more homogeneous cDNA in less time compared to the conventional reverse transcription.
Conventional Reverse Transcription
| Step 1: | RNA denaturation at 65°C for 10 minutes |
| Step 2: | cDNA synthesis at a temperature between 37°C ~ 55°C for 15 ~ 60 minutes |
Cyclic Reverse Transcription
| Step 1: | Primer annealing at a temperature between 15°C and 40°C for 30 seconds |
| Step 2: | cDNA synthesis at a temperature between 42°C and 48°C for 4 minutes |
| Step 3 (optional): | Denaturation of the secondary structure of the RNA template and cDNA synthesis at a temperature between 50°C and 55°C for 30 seconds |
Features and Benefits
| Convenient lyophilized RT premix: | Simply add your purified RNA and start your reaction. Enzyme, dNTPs, reaction buffer, and oligo (dT)20 are provided. |
| Broad range of working temperatures: | For RNAs that are G:C rich or significant secondary structure |
| Novel Cyclic Reverse Transcription: | Ensures that you isolate even the rarest transcripts |
| High yield of cDNA: | For genes up to 9 kb within 10 minutes |
| Stable for 2 years at -20°C: | Long life |
| RNase, DNase and Proteinase-free: | Ensures the integrity of your samples |
Specifications
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment size: 9 kb
Application
cDNA synthesis
Figure 1. Reaction condition comparison of Cyclic Reverse Transcription versus conventional reverse transcription PCR. 800 ng of HeLa cell total RNA was reverse transcribed using different reaction conditions and then 800 bp, 1.5 kb, 2.0 kb, and 2.6 kb of human transferrin receptor gene were amplified using AccuPower PCR PreMix.
M: 1 kb DNA Ladder (D-1040)
Lane 1 - 4: Conventional RT with dT20 at 42°C
Lane 5 – 8: Conventional RT with dN12 at 42°C
Lane 9 - 12: Conventional RT with dT20 at 55°C
Lane 13 - 16: Conventional RT with dN12 at 55°C
Lane 17 - 20: Cyclic RT with dT20 by 55°C
Lane 21 - 24: Cyclic RT with dN12 at 55°C
Figure 2. Coxsackie virus A9 (AJ295200) 5’-UTR region amplification with different reverse transcription products. Coxsackie virus RNA was reverse transcribed with different reverse transcription products and then amplified with PCR with AccuPower PCR PreMix. All materials were used in accordance with manufacturer’s instructions.
M: 100 bp DNA Ladder (D-1030)
Lane 1: Reverse transcription with AccuPower RT PreMix and oligo dT primer
Lane 2: Reverse transcription with AccuPower RT PreMix and random primer
Lane 3 – 4, 7 – 8, 11 – 12: Reverse transcription with AccuPower CycleScript RT PreMix and dT20
Lane 5 – 6, 9 – 10, 13 – 14: Reverse transcription with AccuPower CycleScript RT PreMix and dN12
Lane 15 – 16: Reverse transcription with RT product from company C and oligo dT primer
Lane 17 – 18: Reverse transcription with RT product from company C and random primer
Lane 19 – 20: Reverse transcription with RT product from company I and oligo dT primer
Lane 21 – 22: Reverse transcription with RT product from company I and random primer
Figure 3. Long transcript production comparison. Reverse transcription was carried out with various reverse transcription products prior to PCR with AccuPower PCR PreMix.
M: 100 bp DNA Ladder (D-1030)
Gentaur: Gentaur / 1 hour incubation at 55°C
C: Company C / 1 hour incubation at 42°C
I: Company I / 1 hour incubation at 45°C
Lane 1, 3, 5: Product reverse transcribed with oligo dT primer
Lane 2, 4, 6: Product reverse transcribed with random primer
Figure 4. Transferrin receptor gene amplification with various reverse transcription products. Reverse transcription was carried out with four different reverse transcription products according to manufacturer’s instructions and then cDNA was amplified with AccuPower PCR PreMix.
M: 1 kb DNA Ladder (D-1040)
Lane 1 – 5: Reverse transcribed with AccuPower CycleScript RT PreMix (dT20) / 1 hour at 55°C
Lane 6 – 10: Reverse transcribed with AccuPower CycleScript RT PreMix (dN12) / 1 hour at 55°C
Lane 11 – 15: Reverse transcribed with Company C product and oligo dT primer / 1 hour at 42°C
Lane 16 – 20: Reverse transcribed with Company C product and random primer / 1 hour at 42°C
Lane 21 – 25: Reverse transcribed with Company I product and dT primer / 1 hour at 45°C
Lane 26 – 30: Reverse transcribed with Company I product and random primer / 1 hour at 45°C
Figure 5. Reaction temperatures and time comparison. Data A and B demonstrate high thermal stability of AccuPowerCycleScript RT PreMix.
A: Reverse transcription carried out with AccuPower CycleScript RT PreMix and dT20
B: Reverse transcription carried out with AccuPower CycleScript RT PreMix and dN12
Conventional RT #1: 1 hour incubation at 42°C
Cyclic RT #1: Repeat 12 times of 2 minutes at 37°C and 3 minutes at 50°C
Conventional RT #2: 1 hour incubation at 55°C
Cyclic RT #2: Repeat 12 times of 1 minutes at 37°C, 3 minutes at 47°C, and 3 minutes at 55°C
M: 100 bp DNA Ladder (D-1030)
Lane 1, 5, 9, 13, 17, 21, 25, 29: 100 ng of HeLa cell total RNA used for reaction
Lane 2, 6, 10, 14, 18, 22, 26, 20: 10 ng of HeLa cell total RNA used for reaction
Lane 3, 7, 11, 15, 19, 23, 27, 31: 1 ng of HeLa cell total RNA used for reaction
Lane 4, 8, 12, 16, 20, 24, 28, 32: 100 pg of HeLa cell total RNA used for reaction
Figure 6. Data C and D demonstrate superior productivity of AccuPower CycleScript RT PreMix..
C: Reverse transcription carried out with AccuPower CycleScript RT PreMix and dT20
D: Reverse transcription carried out with AccuPower CycleScript RT PreMix and dN12
10 min: 2 times of 2 minutes at 37°C and 3 minutes at 50°C
20 min: 4 times of 2 minutes at 37°C and 3 minutes at 50°C
60 min: 12 times of 2 minutes at 37°C and 3 minutes at 50°C
M: 100 bp DNA Ladder (D-1030)
Lane 1, 5, 9, 13, 17, 21: 100 ng of HeLa cell total RNA used for reaction
Lane 2, 6, 10, 14, 18, 22: 10 ng of HeLa cell total RNA used for reaction
Lane 3, 7, 11, 15, 19, 2: 1 ng of HeLa cell total RNA used for reaction
Lane 4, 8, 12, 16, 20, 24, 28, 32: 100 pg of HeLa cell total RNA used for reaction
AccuPower Gold Multiplex PCR PreMix
AccuPower Gold Multiplex PCR PreMix can amplify up to 20 target genes in a single tube. AccuPower Gold Multiplex PCR PreMix contains Gentaur’s unique enzyme-mediated HotStart technology with Pyrophosphatase (PPase) and Pyrophosphate (PPi) for efficient suppression of non-specific products and enhanced amplification specificity.
AccuPower Gold Multiplex PCR PreMix can be used for a variety of applications including genotyping assays or molecular diagnostics, and can also be used for cDNA-based semi-quantitative assays.
Features and Benefits
Flexibility: Up to 20 different target genes from human genomic DNA can be amplified in a single tube
Specificity: Pyrophosphate (PPi) has high affinity for Mg2+. By adding PPi to the reaction mixture, the Mg2+ ions necessary for normal PCR are bound, preventing DNA polymerase activity. This PPi-Mg2+ binding prevents non-specific before PCR (zero-cycle) product formation. Upon thermal cycling, the pyrophosphatase (PPase) that is also added to the mixture is activated (>70°C) and hydrolyzes the PPi to 2 phosphate groups and facilitates the release of Mg2+, which is then available for DNA polymerase to use and resume normal activity.
Easy-to-use: All reaction components required for PCR, including thermostable DNA polymerase and dNTPs are contained within each tube and in a lyophilized “PreMix” form. The user needs only to add template DNA, primers and water to perform up to 20-plex PCR. Materials necessary for loading agarose gels for electrophoresis are also added in the reaction, negating the need to add loading dye after PCR is completed.
Reproducibility: Each batch is produced under strict quality controls. Errors that may occur during mass production are eliminated during the individual packaging process. Gentaur’s current batch processing system allows for the production of more accurate and reproducible end-product. Additionally, the streamlining of setup using a lyophilized premix enhances reproducibility by minimizing setup variables.
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 |
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
Each lane from left to right represensts the progressive number of primer sets (1 – 20) included in single AccuPowerGold Multiplex PCR PreMix reactions.
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).
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).
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.
AccuPower TLA PCR PreMix
AccuPower TLA PCR PreMix is a high fidelity PCR master mix that contains an easy to resuspend lyophilized mix of TLADNA Polymerase, dNTPs, reaction buffer, tracking dye, and patented stabilizer conveniently packaged in individual tubes or plates. Simply add template, primers and water. The TLA DNA Polymerase in the premix is a cloned and modified derivative isolated from Thermococcus onnurineus NA1 and features a proofreading activity, an enhanced processivity, and robust amplification efficiency compared to Pfu DNA polymerase or Vent DNA polymerase. These features allow high fidelity DNA amplification as well as long range PCR up to 20 kb.
AccuPower TLA PreMix high fidelity PCR master mix is available with or without loading dye. When loading dye is present you can load it directly into your gel after PCR – no glycerol or glucose is required.
Features and Benefits
| Ease of use: | Just add template and primers and start your PCR, dNTPs, buffer and enzyme are provided |
| Stability: | Stable at room temperature for a month and for 2 years in a -20°C freezer |
| High fidelity: | For subcloning and other critical applications |
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
AccuPower HotStart PCR PreMix
AccuPower HotStart PCR PreMix is a PCR master mix containing a thermostable DNA polymerase, thermostable pyrophosphatase, reaction buffer, dNTPs, tracking dye, and patented stabilizer in a ready to use hotstart PCR master mix.
Gentaur uses a unique enzyme-mediated HotStart PCR that provides robust and reliable results. Gentaur’s Top DNA Polymerase is inhibited at lower temperatures (< 70°C) by pyrophosphate. However, Top DNA Polymerase is rendered fully active at temperatures above 70°C via pyrophosphate hydrolysis with our thermostable pyrophosphatase. This prevents the formation of mis-primed products and primer-dimers during the reaction set up process resulting in improved PCR specificity. It is ideal for nucleic acid amplification reactions involving complex genomic or cDNA templates, very low copy targets, and multiplex reactions.
Features and Benefits
| Ease of use: | Just add template and primers and start your PCR, dNTPs, buffer and enzyme are provided |
| Stability: | Stable at room temperature for a month and for 2 years in a -20°C freezer |
| HotStart: | Unique enzyme mediated HotStart results in greater specificity and more robust reactions |
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)
AccuPower PyroHotStart PCR PreMix
The AccuPower PyroHotStart Taq PCR PreMix is a PCR master mix containing a thermostable DNA polymerase, thermostable pyrophosphatase, reaction buffer, dNTPs, tracking dye, and patented stabilizer in a ready to use hotstart PCR master mix.
Gentaur uses a unique enzyme-mediated HotStart PCR system that provides robust, sensitive and reliable results.
Gentaur's Taq DNA Polymerase is inhibited at lower temperatures (< 70°C) by pyrophosphate. This prevents the formation of mis-primed products and primer-dimers during the reaction set up process resulting in improved PCR specificity. However, Taq DNA Polymerase is rendered fully active at temperatures above 70°C via pyrophosphate hydrolysis with a thermostable pyrophosphatase (Figure 1). AccuPower® PyroHotStart Taq PCR PreMix is ideal for nucleic acid amplification reactions involving complex genomic or cDNA templates, very low copy targets, and multiplex reactions.
Figure 1. The mechanism of an enzyme-mediated hotstart PCR
Features and Benefits
| Ease of use: | Just add template DNA, primers and water to start your reaction. dNTPs, buffer and enzyme are provided. |
| Robust Stability: | Stable at room temperature for a month, at 4°C for one year and for 2 years in a -20°C freezer. |
| Improved specificity: | Unique enzyme mediated PyroHotstart system results in greater specificity and more robust reactions |
| Reproducibility: | Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment. |
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
AccuPower PCR Premix
Gentaur’s AccuPower PCR PreMix is a convenient lyophilized PCR master mix containing Top DNA Polymerase, dNTPs, reaction buffer, tracking dye, and patented stabilizer. AccuPower PreMix PCR master mix includes our super-processive “three times faster than Taq” Top DNA Polymerase for the fastest nucleic acid amplification. While Top DNA Polymerase is ideal for applications where you would normally use Taq, if your needs include HotStart PCR, or High Fidelity PCR, please see our AccuPower HotStart PCR and AccuPower TLA product pages. AccuPower PreMix PCR master mix is available with or without tracking dye, depending on your application. If purchased with tracking dye, reactions can be loaded on agarose gels without adding loading buffer.
Features and Benefits
| Ease of use: | Just add template and primers and start your PCR, dNTPs, buffer and enzyme are provided |
| Stability: | Stable at room temperature for a month and for 2 years in a -20°C freezer |
| Gel loading: | Available with or without a tracking dye for ease of use |
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
AccuPower Taq PCR PreMix
The AccuPower Taq PCR PreMix is a convenient lyophilized PCR master mix containing Taq DNA polymerase, dNTPs, reaction buffer, tracking dye, and patented stabilizer and is aliquoted in 8-strip PCR tubes.
The premix retains its activity for over a month at room temperature and is stable for two years in -20°C freezer.
AccuPower Taq PCR PreMix is available with or without tracking dye, depending on your application. If purchased with tracking dye, reactions can be loaded on agarose gels without adding loading buffer.
Features and Benefits
| Ease of use: | Just add template and primers and start your reaction. All other reaction components including dNTPs, buffer and enzyme are provided. |
| Robust Stability: | Stable at room temperature for a month, at 4°C for one year and for 2 years in a -20°C freezer. |
| Versatility: | The premix is ideal for a wide range of PCR applications. |
| Gel loading: | Available with or without a tracking dye. Samples with tracking dye can be loaded directly into gels after PCR |
| Reproducibility: | Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment. |
Specifications
Source: Thermus aquatics
5' to 3' exonuclease activity: Yes
3' to 5' exonuclease activity: No
3' – A Overhang: Yes
Fragment size: Up to 8 kb from human genomic DNA and 10 kb from Lambda DNA
Applications
- Routine PCR
- Primer extension
- TA cloning
- Gene sequencing
Figure 1. Comparison of PCR amplification efficiency between AccuPower Taq PCR PreMix from Gentaur and other suppliers' PCR master mix.
The cycling conditions for AccuPower Taq PCR PreMix were 95°C for 5 min, 35 cycles of 95°C for 20 sec, 55°C for 20 sec, and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target Gene : Human myc.
Lane M : 100 bp DNA Ladder (Gentaur, Cat. No. D-1030)
Lane 1 : 10 ng of human total cDNA
Lane 2 : 1 ng of human total cDNA
Lane 3 : 100 pg of human total cDNA
Lane 4 : 10 pg of human total cDNA
Figure 2. Comparison of PCR amplification efficiency between AccuPower Taq PCR PreMix from Gentaur and other suppliers' PCR master mix.
The cycling conditions for AccuPower Taq PCR PreMix were 95°C for 5 min, 35 cycles of 95°C for 20 sec, 55°C for 20 sec, and 72°C for 30 sec. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Target Gene: IRGC (Immunity-related GTPase family, cinema)
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 3. Comparison of PCR amplification of long targets between AccuPower Taq PCR PreMix from Gentaur and other suppliers' PCR master mix.
The cycling conditions for AccuPower Taq PCR PreMix were 95°C for 5 min, 30 cycles of 95°C for 20 sec, 65°C for 20 sec, and 68°C for 8 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
Lane M : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
Lane 1 : 3 kb fragment (human tumor protein p53 gene)
Lane 2 : 4 kb fragment (human beta globin region)
Lane 3 : 4.5 kb fragment (human DNA cross-link repair 1A gene)
Lane 4 : 8 kb fragment (human hemoglobin epsilon 1 gene)
PowerChek™ Animal Species ID Real-time PCR Kits
Characteristics
· Detection kit for Animal Species
· Real-time PCR based on TaqMan™ probe-based technology
· Rapid & simple reaction
· No contamination : closed system
· No error : Internal positive control
· Easy to operation in less than 2.5 hrs
Kit Components
· Primer/probe mixture
· 2X Real-time PCR Master Mix
· Control
Protocol
After DNA Ext.
· Step 1 : Mix extracted DNA with the Kit’s reagents
· Step 2 : Real-time PCR run
· Step 3 : Analysis
Exp
Products:
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R0406 |
PowerChekTM Horse species Real-time PCR Kit Fluorephore: FAM |
50 rxn |
383-Eur |
|
R0406E |
PowerChekTM Horse species Real-time PCR Kit with EPC Fluorephore: FAM, HEX(VIC) |
50 rxn |
441-Eur |
New PCR system to discover dangerous foodborne pathogens explored by researcher
Pina Fratamico is on the way to find the easiest and fastest way to test for harmfulEscherichia coli in ground beef. She explores using a next-generation real-time polymerase chain reaction (PCR) system to discover specific gene targets that indicate the presence of dangerous foodborne pathogens. The results show that assays performed using this PCR system are rapid, sensitive, and reliable.
"Testing using these types of systems is faster, easier, and more reproducible than previous methods, and this should increase food safety in the long run. I feel that we could confidently move to these new systems for screening ground beef and other foods for E. coli contamination," says Fratamico, researcher at the USDA Agricultural Research Service in Wyndmoor, Pennsylvania.
Certain strains produce a potentially dangerous toxin called Shiga toxin, but not all E. coli are dangerous. These Shiga toxin-producing E. coli also known as STEC can be found in raw meat and cause serious food poisoning in humans. According the FSIS - Food Safety and Inspection Service website, in October 2012 over, 2,300 pounds of ground beef were recalled due to contamination with STEC.
"Certain groups of STEC have been declared as adulterants by the USDA FSIS, and the availability of rapid and reliable tests for these pathogens is critical so that testing results are available before meat is shipped to restaurants and consumers," she explains.
In the meat industry the PCR protocol has already been used for some time. The genetic test detects the presence of specific gene targets that indicate the existence of STEC in meat. The new generation of real-time PCR systems, like the GeneDisc from France used in this particular study, employ a self-contained unit that standardizes the procedure and tend to be relatively portable and easy to use - offering obvious advantages for both meat processors and inspectors from the industry and government alike.
