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    Displaying items by tag: AccuPower
    Monday, 28 October 2013 18:40

    AccuPower samples available!

    Cat. Product Price Order
    D-1020 SM 25/100 bp mixed DNA ladder 5 € Order
    D-1040 SM 1kb DNA Ladder 5 € Order
    K-2631 SM AccuPower ProFi Taq PCRpremix 5 € Order
    K-2611 SM AccuPower PyroHotstart Taq PCR premix 5 € Order
    K-2301 SM AccuPower Hotstart Pfu peR PreMix 5 € Order
    K-2101 SM AccuPower RocketScript RTPremix 5 € Order
    K-2501 SM AccuPower RocketScript RT-PCR premix 5 € Order
    K-6704 SM AccuPower Dual-HotStart RT-qPCR premix 5 € Order
    K-6251 SM AccuPower 2X GreenStar qPCRMaster Mix 5 € Order
    K-6210 SM AccuPower GreenStar qPCR PreMix 5 € Order
    K-6100 SM AccuPower DualStar qPCR PreMix 5 € Order

     

    More: AccuPower

     

     

     

     

     

    Published in Promos

    CRISPR-mice-virus-detection-teratoma-teratoma-service-ipsc-transgenic-mouseMIT researchers have shown that genes on or off in yeast and human cells by controlling, when DNA into RNA - bypokrok that will allow scientists to better understand the function of these genes copy.
    This technique can also be easier to control the environment of the cell produkujídrogy or disease realize engineer, says Timothy Lu, assistant professor of electrical engineering and computer science and bio - engineering and lead author of a paper describing the new approach in the journal ACS Synthetic Biology.
    "I think it will be much easier to build synthetic circuits" says Lu, a member of the Synthetic Biology Center at MIT. "It should increase the extent and speed with which we can build a number of synthetic circuits in yeast and mammalian cells."
    The new method is based on a system of viral proteins have been used recently for the treatment of genomes of bacteria and human cells. Initial system called CRISPR consists of two parts: a protein which binds to DNA and washers and a short strand of RNA, leaders of the protein into the correct position in the genome.
    "CRISPR system is so powerful that it can be used for a variety of DNA - binding regions of a simple conversion of this handbook RNA targeted basis" says Lu. "By simply reprogramming the RNA sequence of this protein can be any place you want to call the genome or synthetic circuit."
    The main author of the article is Farzadfard Fahim, an MIT graduate student of biology. Samuel Perl, student of Electrical Engineering and Computer Science, is takéautorem.

    Targeting transcription

    In previous studies CRISPR was used to cut pieces of the gene is switched off or replace it with another gene. Lu and his colleagues decided to use the CRISPR system for a different purpose : control of gene transcription is the process by which DNA sequence into RNA (mRNA ), which goes beyond genes copied instructions.
    Transcription is strictly regulated by proteins called transcription factors. These proteins bind to specific DNA sequences in the promoter region of the gene, and either modify or block the enzymes needed for the copy of the gene into mRNA.
    Which acts as a transcription factor for this study, the researchers adjust CRISPR system. Firstly, the modified CRISPR normal protein known as time9 so that it no longer cut the DNA bound thereto. They are also added to the protein, the segment that is activated or suppressed by modulating gene expression machinery of cellular transcription.
    Time9 to get to the right place, the researchers also corresponds delivered to the target cells, the genes for RNA guides who chtějísekvence DNA activates the promoter of the gene.
    Researchers have shown that when the guide RNA and protein time9 connection in the target cell, the target gene přesněpravý atranskripce. To their surprise, the same complex time9 also used to block transcription to be found in other parts of the gene.
    "It's nice that allows you to make a positive and negative regulation of the same protein, but with different RNA targeted to various managerial positions in the promoter" says Lu.

    "A lot of elasticity"

    The new system should be much simpler than other newly developed two systems for the transcription of DNA - binding proteins such as zinc finger transcriptional activator and effector Nucleases ( Talens ) is known, says Lu. Although effective, the construction and assembly of proteins is time- consuming and expensive.
    "There is a lot of flexibility with CRISPR, and it really comes from the fact that you do not spend more time in protein engineering. You can only change the sequence of the nucleic acid RNA, " says Lu.
    "The fact that this can be used for effective regulation of transcription in both yeast and mammalian cells, it is very encouraging," says Kobi Benenson, Professor of Biosystems Science and Engineering at ETH Zurich, which is not part of the research team." This technology may be used in the very near future genetic engineering and synthetic biology applications for biopharmaceuticals - "Tissue engineering and gene therapy, among other things"
    Researchers also transcriptional control designed so that it can be triggered by specific small molecules, which may be included in the cell, such as sugars. For this, the guide RNA genes constructed so that they are only produced Primal molecule is available. No small molecules, there are no guidelines aRNA gene is targeted at rest.
    This type of control could be used to explore the role of naturally occurring genes on and off at specific times during development or progression useful, says Lu.
    Lu is now working on the development of advanced synthetic circuits for applications such decisions are based on multiple inputs made ​​by the mobile environment. " We want to be able to scale- up the most complex circuits and show that anyone is ever built in yeast and mammalian cells" he says.

    Published in News
    Thursday, 30 May 2013 09:58

    AccuLadder 100 bp DNA Size Marker

    More Sharp! High Resolution! More Convenience!

    AccuLadder 100 bp DNA size marker was designed to determine the size of double stranded DNA fragments from 100 to 2,000 bp. AccuLadder is shaper and brighter than our standard 100bp ladder The AccuLadder 100 bp DNA size marker consists of 13 double stranded molecular weight markers ranging in sizes from 100 to 1,000 bp in 100 bp increments, and additional fragments of 1,200, 1,600, 2,000 bp. The 500, 1,000 and 2,000 bp bands are two times brighter for easy identification.

    Note:
    The DNA ladder can be applied directly onto an agarose gel.
    There is no need to heat before loading. Repeated freezing and thawing should be avoided.

    Features and Benefits

    Complete and ready to load: Convenient
    Easy-to-Identify reference bands: Easy-to-read results
    Competitive pricing: Great value for your research dollar

     

    Specifications

    Concentration: 54 ng/μl
    Recommended loading: 4.0~5.0 μl / 5 mm lane width
    Typical Number of lanes: 126~157 (5 mm lane width)
    Size range (bp): 100 - 2,000
    Number of bands: 13
    Supplied in: 10 mM Tris-HCl (pH 8.0), 1 mM EDTA, 2.5% Ficoll, 0.005% Bromphenol Blue, 0.005% Xylene Cyanol
    Storage: -20°C

    acculadder100bpDNA figure1

    2.0% TBE agarose gel stained with Ethidium Bromide.

    Nuclease Activity Test

    acculadder100bpDNA figure2

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    Published in Promos
    Wednesday, 29 May 2013 16:18

    AccuPower ProFi Taq PCR PreMix

    AccuPower® ProFi Taq PCR PreMix for high efficiency and amplification of long range PCR.

    ProFiTaqPCRPreMix f01

    AccuPower® ProFi Taq PCR PreMix is a convenient lyophilized PCR master mix containing ProFi Taq DNA polymerase, reaction buffer, dNTPs, tracking dye, and a patented stabilizer. ProFi Taq DNA polymerase in the premix is a unique recombinant Taq DNA polymerase that offers enhanced amplification efficiency and higher fidelity for PCR. AccuPower® ProFi Taq PCR PreMix is applicable to any template DNA, and especially effective in amplifying large genomic DNA fragments around 20 kb. AccuPower® ProFi Taq PCR PreMix provides accurate long-range amplification of standard and amplification of low-copy target, and is highly suitable for all PCR applications.

     

    Features and Benefits

    Long PCR: ProFi Taq is especially effective in amplifying large genomic DNA fragments around 20 kb and amplifying Lambda DNA up to 30kb.
    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.
    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 and for 2 years in a -20°C freezer

     

    Specifications

    5' to 3' exonuclease: Yes
    3' to 5' exonuclease: Yes
    3' – A Overhang: Yes
    PCR product size: ~ 30kb


    Application

    - Primer extension
    - long-range amplification from genomic DNA
    - High amplification efficiency
    - Excellent performance on difficult templates
    - Amplification of low-copy targets
    - High yield and high sensitivity PCR

    Experimental data

    ProFi Taq fig1

    Figure 1. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 30 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 : human insulin receptor gene.
    Lane M : 100 bp DNA Ladder(Gentaur, Cat. No. D-1030)
    Lane 1 : 10 ng of human genomic DNA
    Lane 2 : 1 ng of human genomic DNA
    Lane 3 : 100 pg of human genomic DNA
    Lane 4 : 10 pg of human genomic DNA

    ProFi Taq fig2
    Figure 2. Comparison of PCR amplification efficiency between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' 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) wase used as a template for PCR amplification. The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5min, 33 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 : human GAPDH gene.
    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

    ProFi Taq fig3
    Figure 3. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi 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 4 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol.
    Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
    Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
    Lane 1 : 2 kb fragment (human tumor protein p53 gene)
    Lane 2 : 3 kb fragment (human tumor protein p53 gene)
    Lane 3 : 4.5kb fragment (human DNA cross-link repair 1A gene)
    Lane 4 : 8 kb fragment (human hemoglobin epsilon 1 gene)

    ProFi Taq fig4
    Figure 4. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec and 68°C for 15 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Human genomic DNA was used as a template for PCR amplification.
    Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
    Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
    Lane 1 : 11 kb fragment
    Lane 2 : 13.5 kb fragment
    Lane 3 : 17.6 kb fragment
    Lane 4 : 21.4 kb fragment

    ProFi Taq fig5
    Figure 5. Comparison of PCR amplification of long targets between AccuPower® ProFi Taq PCR PreMix from Gentaur and other suppliers' PCR master mix

    The cycling conditions for AccuPower® ProFi Taq PCR PreMix were 95°C for 5 min, 32 cycles of 95°C for 20 sec, 65°C for40 sec, and 68°C for 20 min. PCR reactions using other suppliers' PCR master mix were performed according to each supplier's protocol. Lambda DNA was used as a template for PCR amplification.
    Lane M1 : Lambda/Hind III marker (Gentaur, Cat. No. D-1050)
    Lane M2 : 1 kb DNA Ladder (Gentaur, Cat. No. D-1040)
    Lane 1 : 15 kb fragment
    Lane 2 : 20 kb fragment
    Lane 3 : 25 kb fragment
    Lane 4 : 30 kb fragment

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    Published in Promos

    Ready-to-use cocktail containing all components,except primer, for the amplification and detection of DNA in real-time quantitative PCR(qPCR).

    The AccuPower® 2X Greenstar qPCR Master Mix is a ready-to-use cocktail containing all components,except primer, for the amplification and detection of DNA in real-time quantitative PCR(qPCR). It combines the automatic "Hotstart" technology of Top DNA polymerase and SYBR Green I fluorescent dye to deliver excellent sensitivity in the quantification of target sequences, with a linear dose response over a wide range of target concentration. Volumes are provided for 100 or 200 amplification reactions of 50ul each.

    accupower-greenstarmaster product f01

    Features and Benefits

    High Specificity : AccuPower® 2X Greenstar qPCR Master Mix provides more accurate Real-time PCR result by application of Hot-start method.
    Stability: The chemical stabilizer maintains enzyme activity for 2 years at -20°C
    Simplicity: Ready to use, AccuPower® 2X Greenstar qPCR Master Mix contains everything of Real-time PCR excluding primer and template.
    Reproducibility: Gentaur's strict quality controlled production system ensures that your results will be reproducible experiment after experiment

     

    Applications

    - Real-time quantification of DNA and cDNA targets
    - Gene expression profiling
    - Microbial & Viral pathogen detection

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    Published in Promos
    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 12:15

    AccuPower RT-PCR PreMix

    AccuPower RT-PCR PreMix contains all the components necessary for sequential cDNA synthesis and amplification in one tube (one-step RT-PCR). This RT-PCR PreMix consists of both M-MLV Reverse Transcriptase, RNA dependent DNA polymerase, and a thermostable DNA polymerase in a lyophilized mix of dNTPs, reaction buffer, RNase inhibitor, tracking dye, and stabilizer. The kit can be used for double stranded cDNA synthesis from low copy RNA or mRNA and subsequent RT-PCR .

     

    Features and Benefits

    Convenient lyophilized RT: Easy to use, simply add your purified RNA
    High yield of cDNA: For genes up to 6 kb
    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: 6 kb


    Application

    cDNA synthesis followed by PCR

    14CycleScript RT gentaur RT-PCR PreMix RocketScript gentaur Cycle RT antibodies PreMix

    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

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