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GENTAUR Europe

 GENTAUR Europe BVBA
Voortstraat 49, 1910 Kampenhout BELGIUM
Tel 0032 16 58 90 45 
Fax 0032 16 50 90 45
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 GENTAUR BULGARIA
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    Gentaur London

     GENTAUR Ltd. 
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    1404-1410 High Road 
    Whetstone London N20 9BH 
    Tel 020 3393 8531 
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     GENTAUR Poland Sp. z o.o. 

    ul. Grunwaldzka 88/A m.2

    81-771 Sopot, Poland
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    Tel 0208-080893 
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    Piazza Giacomo Matteotti, 6, 24122 Bergamo
    Tel 02 36 00 65 93 
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    Other countries

    Other countries
    Luxembourg +35220880274
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    Friday, 01 March 2013 13:45

    AccuPower DNA Ligation PreMix

    18AccuPowerDNALigationantibodies PreMix DualStar qPCR

    The AccuPower DNA Ligation PreMix is a lyophilized master mix containing T4 DNA Ligase, ATP, reaction buffer, and patented stabilizer. This DNA ligation premix is conveniently aliquoted in strip-tubes for reactions; you need only add DNAs to be ligated and water. The reaction will work for DNA ligation for all applications: blunt cloning, sticky end cloning and TA cloning. The premix is stable up to four months at room temperature and for three years at -20°C.

     

    Features and Benefits

    Ready to use premix: Minimal set up time and handling required
    Fast: Only 5 minutes for cohesive-end ligation and 10 minutes for blunt-end ligation at room temperature
    Stable: Enzyme activity for up to four months at room temperature and for three years in the freezer



    Application

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

    19taq PyroHotStart HotStart

    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

    20pcr gentaur taq PyroHotStart HotStart

    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

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    Published in Promos
    Friday, 01 March 2013 13:33

    AccuPower DualStar qPCR PreMix

    AccuPower DualStar qPCR PreMix is a lyophilized PCR master mix containing a thermostable DNA polymerase, thermostable pyrophosphatase, reaction buffer, dNTPs, tracking dye, and patented stabilizer in a ready to use hotstart qPCR master mix.

    DualStar qPCR PreMix eliminates nonspecific reaction, while providing high sensitivity, due to our novel HotStart methodology. Gentaur uses a unique enzyme-mediated HotStart PCR that provides robust and reproducible 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.

    DualStar Plus qPCR PreMix goes a step further by providing resistance to common inhibitors of PCR (Blood-EDTA, Hemoglobin, Humic Acid from various sources, etc.) providing you with robust and reliable results, even with poor quality samples. Tested for resistance against many common inhibitors of PCR, DualStar plus is the enzyme of choice for any PCR that you want to work right – the first time and every time!

     

    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
    Reproducibility: Each batch is produced under strict ISO quality controls.
    DualStar Plus qPCR PreMix: Available for less samples that will not work in other systems due to low purity

    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

    15antibodies premix accupower RocketScript

    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).

    16elisa pcr gentaur taq PyroHotStart HotStart

    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).

    17RT gentaur RT-PCR PreMix RocketScript gentaur Cycle RT

    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).

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    Published in Promos
    Friday, 01 March 2013 13:18

    AccuPower RocketScript Cycle RT PreMix

    AccuPower RocketScript Cycle RT PreMix is a ready-to-use lyophilized mastermix containing all components for first-strand cDNA synthesis from purified Poly(A) or total RNA template. 
    The premix contains Gentaur's exclusive M-MLV based thermostable reverse transcriptase (RTase), RocketScript, and oligo dT20 for added convenience. Conditions are optimized for Gentaur's patented Cyclic Temperature Reverse Transcription (CTRT) in a premix form.

    Native M-MLV RTase has maximum activity at relatively low temperatures (42°C), causing several problems in reverse 
    transcription of RNA molecules with complex secondary structure. AccuPower RocketScript Cycle RT PreMix has thermostable activity across a wide temperature range, (42°C – 70°C), allowing efficient cDNA synthesis from virtually any RNA. 
    The lyophilized PreMix contains all components necessary for a successful CTRT reaction, including RTase, RNase inhibitor and buffer components. Just add template RNA, primers and water, and the RT reaction is ready to go.

    accupower-antibodies-gentaur-aproketscript overview 

    Schematic representation of the 5' UTR of a gene, with complex secondary structure, at three different temperatures. Note that RocketScript shows full activity at 70°C allowing it to synthesize the complete gene sequence where M-MLV and other Reverse Transcriptase's fail.

    Features and Benefits

    Thermostable Activity: RocketScript is able to perform reverse transcription reactions throughout a wide range 
    of temperatures from 42°C to 70°C. 

    Enhanced Performance: RocketScript has enhanced performance to handle high and low input RNA concentrations 
    as well as short and long RT target sizes. 

    Easy-to-use: The product contains the enzyme itself, plus RNase inhibitors and all other components necessary for the best reverse transcription results in the tube. 

    Reproducibility: Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment. 

    Flexible Reaction Conditions: Both CTRT and first-strand cDNA synthesis are both possible, with reaction temperatures 
    ranging from 42°C to 70°C. 

    Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided. 

    Stability: Stable at room temperature for a month one year at 4°C and for 2 years in a -20°C freezer 

    RNase, DNase and Proteinase-free: Ensures the integrity of your samples. 

    Broad range of working temperatures: For RNAs that are G:C rich or significant secondary structure 

    Stable for 2 years at -20°C: Long shelf life 

    Specifications

    5' to 3' exonuclease: No
    3' to 5' exonuclease: No
    3’ – A Overhang: No
    Fragment Size: Up to 10 kb


    Application

    - First-strand synthesis of cDNA from RNA molecules (Reverse Transcription)
    - RT-PCR
    - Random priming reaction
    - Library construction
    - Probe labeling
    - mRNA 5’end mapping by primer extension analysis
    - Real time PCR


    Experimental data

    accupower-antibodies-gentaur-aproketscript-cycle figure1

    Figure 1. Comparison of amplification efficiency between AccuPower RocketScript Cycle RT PreMix and competitors’ RTases
    (a) Sensitivity test
    Primer set: Human TFRC set
    Lane M: 1 kb DNA Ladder
    Lane1: 100 ng Human total RNA from HeLa cell
    Lane 2: 10 ng Human total RNA from HeLa cell
    Lane3: 1 ng Human total RNA from HeLa cell
    Lane4: 100 pg Human total RNA from HeLa cell
    RT reaction condition is performed according to each manufacturer’s recommendations

    (b) Full-Length cDNA synthesis test
    RT reactions were performed according to each manufacturer’s recommendation. All cDNAs were amplified with AccuPower Hotstart PCR Premix (K-5050) from Gentaur
    Note: Competitor products show inhibition at high input concentrations of total RNA
    Lane 1: 1 μg Human total RNA from HeLa cell
    Lane 2: 100 ng Human total RNA from HeLa cell
    Lane 3: 10 ng Human total RNA from HeLa cell

    accupower-antibodies-gentaur-aproketscript-cycle figure2

    Figure 2. Complex RNA amplification results of RocketScript Cycle RT PreMix
    Each target gene was amplified after performing reverse transcription with AccuPower RocketScript Cycle RT PreMix.
    Reverse transcription conditions: Conventional 1 hr incubation at 42°C, 50°C, or 60°C, deactivation at 95°C for 5 min
    A: M-MLV Reverse Transcriptase
    B: AccuPower RocketScript Cycle RT PreMix with Oligo (dT)20
    Lane 1: 100 ng Human total RNA from HeLa cell
    Lane 2: 10 ng Human total RNA from HeLa cell

     

    Concentration (copies/rxn) FTRT (Ct) CTRT with 1 cycle (Ct) CTRT with 10 cycles (Ct)
    10,000 19.46 18.77 18.51
    1,000 24.11 24.04 22.93
    100 29.78 28.35 28.19
    10 32.87 33.00 31.05

     

     accupower-antibodies-gentaur-aproketscript-cycle figure3

    Figure 3. Low copy species enrichment by cycle.
    Comparing FTRT (Fixed Temperature Reverse Transcription) to 1 and 10 cycle(s) of CTRT reveal progressive improvement in detected cDNA yield as input copies decrease.

    FTRT: 60 min incubation at 50°C followed by 5 min deactivation at 95°C
    CTRT: Cycles of 37°C annealing 10 sec, 50°C cDNA synthesis 4 min, 55°C secondary structure melting and cDNA synthesis 
    30 sec
    Primer set: Human GAPDH
    Human Total RNA from HeLa cells
    qPCR with AccuPower GreenStar qPCR PreMix (K-6210)

     accupower-antibodies-gentaur-aproketscript-cycle figure4

    Figure 4. Amplification comparison by cycle 

    FTRT (Fixed Temperature Reverse Transcription) conditions:

     

    Step Temperature Time No. of Cycles
    dT20
    cDNA synthesis 50°C 60 min 1
    Heat inactivation 95°C 5 min 1



    CTRT (Cyclic Temperature Reverse Transcription) conditions:

     

    Step Time Temperature No. of Cycles
    dT20
    Primer annealing 37°C 10~30 sec 1, 5, 10, or 15
    cDNA synthesis 50°C 4 min
    Melting secondary structure & cDNA synthesis 55°C 30 sec
    Heat inactivation 95°C 5 min 1

     

    Primer set: Human GAPDH and myc set
    Lane M: 1 kb DNA Ladder
    Lane1: 10 ng Human Total RNA from HeLa Cell
    Lane 2: 1 ng Human Total RNA from HeLa Cell
    Lane 3: 100 pg Human Total RNA from HeLa Cell
    Lane 4: 10 pg Human Total RNA from HeLa Cell

    accupower-antibodies-gentaur-aproketscript-cycle figure5

    Figure 5. Comparison of amplification results between Gentaur RocketScriptTM Cycle RT PreMix and competitor RTases
    All cDNAs were amplified with AccuPower Hotstart PCR Premix (K-5050) from Gentaur
    Primer set: human Myc498 bp set
    Lane M: 1 kb DNA Ladder
    Lane1: 10 ng Human total RNA from HeLa Cell
    Lane 2: 1 ng Human total RNA from HeLa Cell
    Lane 3: 100 pg Human total RNA from HeLa Cell
    Lane 4: 10 pg Human total RNA from HeLa Cell

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    Published in Promos
    Friday, 01 March 2013 13:06

    AccuPower RocketScript RT PreMix

    AccuPower RocketScript RT PreMix contains Gentaur’s exclusive M-MLV based thermostable reverse transcriptase (RTase), RocketScript. Native M-MLV RTase has maximum activity at relatively low temperatures (42°C), causing several problems in reverse transcription of RNA molecules with complex secondary structure. 

    RocketScript has thermostable activity (42°C~70°C), allowing efficient cDNA synthesis from virtually any RNA. The lyophilized PreMix contains all components necessary for a successful reverse transcription reaction, including RTase, RNase inhibitor and buffer components. Just add template RNA, primers and water, and the RT reaction is ready to go.

    accupower-gentaur-antibodies-aproketscript overview

     

    Schematic representation of the 5’UTR of a gene, with complex secondary structure, at three different temperatures. Note that RocketScript shows full activity at 70°C allowing it to synthesize the complete gene sequence where M-MLV and other Reverse Transcriptase's fail. 

    Features and Benefits 

    Thermostable Activity: RocketScript is able to perform reverse transcription reactions throughout a wide range 
    of temperatures from 42°C to 70°C. 

    Enhanced Performance: RocketScript has enhanced performance to handle both high and low input RNA 
    concentrations as well as short and long RT target sizes. 

    Easy-to-use: The product contains the enzyme itself, plus RNase inhibitors and all other components necessary for the best reverse transcription results in the tube. 

    Reproducibility: Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment. 

    Convenient: Just add template and primers and start your reaction. dNTPs, buffer and enzyme are provided. 

    Stability: Stable at room temperature for a month one year at 4°C and for 2 years in a -20°C freezer 

    RNase, DNase and Proteinase-free: Ensures the integrity of your samples. 

    Broad range of working temperatures: For RNAs that are G:C rich or significant secondary structure 

    Stable for 2 years at -20°C: Long shelf life 

    Specifications

    5' to 3' exonuclease: No
    3' to 5' exonuclease: No
    3’ – A Overhang: No
    Fragment Size: Up to 10 kb


    Application

    - First-strand synthesis of cDNA from RNA molecules (RT)
    - RT-PCR
    - Random priming reaction
    - Library construction
    - Probe labeling
    - mRNA 5end Mapping by Primer Extension Analysis
    - Real time PCR

    accupower-gentaur-antibodies-aproketscript figure1

    Figure 1. Sensitivity comparison between AccuPower RocketScript RT PreMix and M-MLV RTase
    Sensitivity results of AccuPower RocketScript RT PreMix using GAPDH compared with conventional Reverse transcriptases. 
    Each 100 ng – 10 fg of total RNA used for RT and the same amount of RT products used for electrophoresis.
    Lane 1: 10 fg Human total RNA from HeLa cell           Lane 2: 100 fg Human total RNA from HeLa cell
    Lane 3: 1 pg Human total RNA from HeLa cell            Lane 4: 10 pg Human total RNA from HeLa cell
    Lane 5: 100 pg Human total RNA from HeLa cell        Lane 6: 1 ng Human total RNA from HeLa cell
    Lane 7: 10 ng Human total RNA from HeLa cell          Lane 8: 100 ng Human total RNA from HeLa cell

    accupower-gentaur-antibodies-aproketscript figure2

    Figure 2. Comparison of amplification efficiency between AccuPower RocketScript RT PreMix and competitorsM-MLV RTase.
    RocketScript is able to handle a wide range of sample concentrations and transcript lengths so your downstream applications are minimally effected by the reverse transcription step.
    Lanes 1-3: 1,000 ng, 100 ng and 10 ng of total RNA from HeLa cells, respectively.

    accupower-gentaur-antibodies-aproketscript figure3

    Figure 3. Sensitivity comparison between AccuPower RocketScript RT PreMix and competitor RTases using Real Time PCR
    Reverse transcription conditions: conventional 1 hr incubation at 60°C, deactivation at 95°C for 5 min All cDNAs were amplified with AccuPower DualStar™ qPCR PreMix (K-6110) from Gentaur.

    Concentration RocketScript RT PreMix Supplier Q Supplier I
    10,000 23.91 25.63 24.43
    1,000 27.33 28.92 28.03
    100 30.62 32.42 30.88
    10 33.63 35.43 33.95
    Efficiency 104% 103% 108%
    Linearity 0.99999 0.9996 0.9995

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    Published in Promos
    Friday, 01 March 2013 11:48

    AccuPower RT PreMix

    The AccuPower RT PreMix is a master mix for cDNA synthesis that consists of an easy to resuspend, lyophilized mix ofM-MLV Reverse Transcriptase, RNase inhibitor, dNTPs, reaction buffer, tracking dye, and patented stabilizer. The master mix kit is used for first strand cDNA synthesis from RNA. Downstream applications include cDNA amplification with reverse transcription PCR. All of the key components are premixed at optimal concentrations. Simply add template RNA, primers and water to start your reaction.

    For cDNA amplification with Cyclic RT, please see our AccuPower CycleScript RT PreMix.

     

    Features and Benefits

    Convenient lyophilized RT: Easy to use, simply add your purified RNA
    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

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


    Application

    cDNA synthesis

    13elisa pcr gentaur taq PyroHotStart HotStart

    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

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    Published in Promos
    Friday, 01 March 2013 11:39

    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

    accupower-gentaur-antibodies-cyclescript-figure1

    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

    accupower-gentaur-antibodies-cyclescript-figure2

    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

    accupower-gentaur-antibodies-cyclescript-figure3

    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

    accupower-gentaur-antibodies-cyclescript-figure4

    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

    accupower-gentaur-antibodies-cyclescript-figure5

    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

    accupower-gentaur-antibodies-cyclescript-figure6

    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

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    Friday, 01 March 2013 10:48

    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

    3RocketScript gentaur Cycle RT antibodies PreMix DualStar qPCR gentaur antibodies PreMix DNA

    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

    4PreMix RocketScript gentaur Cycle RT antibodies PreMix

    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)

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    Published in Promos
    Friday, 01 March 2013 10:49

    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.

    accupower-gentaur-antibodies-Enzyme-mediated-HotStart 

    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

    accupower-PyroHotstart Taq antibodies-gentaur-1

    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)

    accupower-gentaur-antibodies-PyroHotstart Taq fig2

    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

    accupower-PyroHotstart Taq -gentaur-antibodies-fig3

    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)

    accupower-gentaur-antibodies-PyroHotstart Taq fig4

    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

    accupower-antibodies-gentaur-PyroHotstart Taq fig5

    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

     

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    Published in Promos
    Friday, 01 March 2013 10:17

    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

    accupower-TaqPCR-gentaur-antibodies-cell-culture-pcr

    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

    accupower-TaqPCR-gentaur-antibodies-cell-culture-pcr-2

    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

    accupower-TaqPCR-gentaur-antibodies-cell-culture-pcr-3

    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)

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    Published in Promos