An optimized ready-to-use solution containing Hot Start Taq DNA polymerase, standard Hot Start Taq reaction buffer, dNTPs, tracking dye and stabilizers. It is ideally suited to routine PCR applications such as sub-cloning, colony screening and genotyping. It can amplify up to 6 kb from complex genomic DNA. Blue dye migrates approximately at 4 kb. 10x1 ml. Storage: -20ºC ***** This is a cold item. You are requires to select Overnight shipping for purchase this item! *****

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Ready to use solutions optimized for Probe real time PCR (Taqman, Molecular Beacon etc) Include the HotStar DNA Polymerase, dNTPs, ROX and the stabilizing agent and an optimized PCR buffer, ensuring PCR specifity and sensitivity. The ROX in the mix is used for normalization of the flurescent signal in the qPCR. Compatible with real time thermal cyclers for probe real time PCR.

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Ready to use solutions optimized for Probe real time PCR (Taqman, Molecular Beacon etc) Include the HotStar DNA Polymerase, dNTPs, ROX and the stabilizing agent and an optimized PCR buffer, ensuring PCR specifity and sensitivity. The ROX in the mix is used for normalization of the flurescent signal in the qPCR. Compatible with real time thermal cyclers for probe real time PCR.

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Ready to use solutions optimized for Probe real time PCR (Taqman, Molecular Beacon etc) Include the HotStar DNA Polymerase, dNTPs, ROX and the stabilizing agent and an optimized PCR buffer, ensuring PCR specifity and sensitivity. The ROX in the mix is used for normalization of the flurescent signal in the qPCR. Compatible with real time thermal cyclers for probe real time PCR.

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2x Taq PCR Premix is an optimized ready-to-use solution containing Taq DNA polymerase, standard Taq reaction buffer, dNTPs, tracking dyes and stabilizers. It is ideally suited to routine PCR applications such as sub-cloning, colony screening and genotyping. It can amplify up to 4 kb from complex genomic DNA or up to 5 kb from lambda DNA. Tracking dyes included in the mix: Blue dye migrates approximately 4 kb, and yellow dye migrates approximately 50 bp. ***** This is a cold item. You are requires to select Overnight shipping for purchase this item! *****

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2x Taq PCR Premix is an optimized ready-to-use solution containing Taq DNA polymerase, standard Taq reaction buffer, dNTPs and stabilizers. It is ideally suited to routine PCR applications such as sub-cloning, colony screening and genotyping. It can amplify up to 4 kb from complex genomic DNA or up to 5 kb from lambda DNA.  No tracking dye included in the mix. 20x1ml, contains 1000U Taq DNA polymerase

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Tris-Glycine Native Sample Buffer (2x) is used to prepare protein samples for native gel electrophoresis using Tris-Glycine gels or Tris Acetate gels. It has a pH of 6.8 and contains bromophenol blue as a tracking dye. To use: Heat the sample in a 1x dilution (reduced or non-reduced) at 85°C for 2–5 minutes for optimal results.Composition: 200 mM Tris HCl, 20% Glycerol, 0.01% Bromophenol blue (Non-reduced). Add ß-mercaptoethanol (8% V/V) before use (Reduced).   MSDS

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Tris-Glycine Native Sample Buffer (2x) is used to prepare protein samples for native gel electrophoresis using Tris-Glycine gels or Tris Acetate gels. It has a pH of 6.8 and contains bromophenol blue as a tracking dye. To use: Heat the sample in a 1x dilution (reduced or non-reduced) at 85°C for 2–5 minutes for optimal results.Composition: 200 mM Tris HCl, 20% Glycerol, 0.01% Bromophenol blue (Non-reduced). Add ß-mercaptoethanol (8% V/V) before use (Reduced).   MSDS

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Tris-Glycine SDS Sample Buffer (2X) is used to prepare protein samples for denaturing gel electrophoresis using Tris-Glycine gels. It has a pH of 6.8 and contains bromophenol blue as a tracking dye. To use: Heat the sample in a 1X dilution (reduced or non-reduced) at 85°C for 2–5 minutes for optimal results. Note: Heating samples at 100°C in SDS-containing buffers results in proteolysis. Composition: 125 mM Tris HCl, 4% SDS, 20% Glycerol, 0.01% Bromophenol blue (Non-reduced). Add ß-mercaptoethanol (8% V/V) before use (Reduced).   MSDS

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Tris-Glycine SDS Sample Buffer (2X) is used to prepare protein samples for denaturing gel electrophoresis using Tris-Glycine gels. It has a pH of 6.8 and contains bromophenol blue as a tracking dye. To use: Heat the sample in a 1X dilution (reduced or non-reduced) at 85°C for 2–5 minutes for optimal results. Note: Heating samples at 100°C in SDS-containing buffers results in proteolysis. Composition: 125 mM Tris HCl, 4% SDS, 20% Glycerol, 0.01% Bromophenol blue (Non-reduced). Add ß-mercaptoethanol (8% V/V) before use (Reduced).   MSDS

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Features The kit is designed for multiplex qPCR with TaqMan probes. The kit uses Super Taq-Probe DNA polymerase specially engineered for TaqMan probes, which increases 5’ to 3’ exonuclease efficiency and produces S-shaped curve. Up to four pairs of gene-specific primers can be applied in one reaction. The concentrations of the primers and probes are variable depending on specific assays and thermocycling protocols (Table 1).  The preferable PCR product size is ≤150bp. The kit has three formulations of No ROX, Low ROX or High ROX concentrations for your choice (see Table 2). Intended Use The 2x Multiplex qPCR Master Mix is used for real-time qualitative and quantitative multiplex qPCR with TaqMan probes for up to four targets.  The master mix is a premixed, 2X concentrated solution that has all the components except for gene-specific primers, probes and templates. Transportation and storage The kit can be transported at below 4°C for up to 3 days.  The kit should be kept stable in the dark at -20°C for ≤24 months with ≤10 times of freeze-thaw cycles. The kit can be stored at 4°C for a week.  Setup Reaction and Thermocycling  Thaw 2x Master mix and other reaction components at room temperature, mix each component, centrifuge and then place on ice.    Set up reactions (Table 1).  Seal tubes with flat, optically transparent caps or seal plate with optically transparent film. Mix and then briefly centrifuge the tubes or plate. Program PCR instrument with indicated thermo-cycling protocol. Load PCR tubes or plate and start to run. Perform data analysis according to the PCR instrument instructions. Table 1. Setting up a 20μl or 10μl Reaction Component Volume per 20μl Volume per 10μl Final Concentration 2x Mastermix 10μL 5μL 1X Primersa Variable Variable Each 150-400nM TaqMan probesb Variable Variable Each 150-250nM DNA Templatec Variable Variable ≤500ng human genomic DNA/20μL H2O To 20μL To 10μL Note: Each primer’s Tm should be designed ≥60°C, preferably between 62°C to 65°C, using primer3 software for high efficiency and specificity. Each probe’s Tm should be 8-10°C higher than the primer’s Tm, preferably between 70-75°C. DNA templates should be extracted by a qualified silica-based kit and eluted with low EDTA TE buffer (10mM Tris-HCl, 0.1mM EDTA, pH 8.0-8.3). Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 95°C 2 min 1 II 95°C 10 sec 35-40 II 60°C, signal acquisition 60 sec 35-40 Table 3. Three-Step Thermocycling Protocol Stage Temperature Period Number of Cycles I 95°C 1 min 1 II 95°C 10 sec 35-40 II 60°C 30 sec 35-40 II 68-72°C, signal acquisition 30 sec 35-40 Notes: The three-step thermocycling protocol in Table 4 increases overall polymerase activity by 50%, a more effective protocol than Table 3. The primer concentration used in Tables 3 and 4 is typically 0.15-0.2uM. Precautions If you order a “No ROX” master mix but you have an Applied Biosystems or Thermo Fisher instrument, please turn off ROX passive reference dye button when setup assays. Table 4. Compatible Instruments qPCR Instrument ROX required by instrument Passive dye setup Bio-Rad® iQ™5, CFX96, CFX384, Opticon Roche Lightcycler®Qiagen Rotor-Gene™Eppendorf Mastercycler®Cepheid® SmartCycler® Not recommended Not necessary Applied Biosystems® 7500, 7500 Fast, QuantStudio™, ViiA7™, Agilent Mx™ Low ROX(50nM final concentration) Turn on ROX passive reference dye  Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne™, StepOnePlus™ High ROX(500nM final concentration) Turn on ROX passive reference dye 

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SYBR Green I-based qPCR reagent: It contains PowerPCR™ Hot Start Taq DNA polymerase, PCR buffer, dNTPs, SYBR Green I fluorescent dyes, Mg2+ and ROX calibration dye. It can be applied to the target sequence detection of genomic DNA and cDNA. The SYBR Green I dye binds to dsDNA without using sequence-specific probes. The chemically modified PowerPCR™ Hot Start Taq DNA polymerase is inactive at room temperature which effectively avoids the non-specific amplification caused by primer dimers or the nonspecific binding of prime and template. The enzyme should be activated by incubation at 95°C for 10 minutes. The unique PCR buffer components and the Hot start enzyme ensure PCR specificity and sensitivity. The ROX is a dye molecule that can be used to normalize the well-to-well fluorescent fluctuations and can be applied to all qPCR instruments using ROX as calibration dye. It has a higher sensitivity and a wider linear range which can exactly quantitate the target gene or low copy template. Suitable Real-Time PCR detection systems: ABI Prism7000/7300/7700/7900 ABI Step One/StepOne Plus Eppendorf systems need high ROX signals

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