| Description | Product content F665904Component500 U2500 UStorageF665904AFastStar DNA Polymerase, 5 U/µL 100 µL5×100 µL-20℃. Avoid freeze/thaw cycle.F665904B10×PCR Buffer 1.8 mL5×1.8 mL-20℃. Avoid freeze/thaw cycle.Notes: 10×PCR Buffer contains 15mM Mg2+. Product Product content F665904Component500 U2500 UStorageF665904AFastStar DNA Polymerase, 5 U/µL 100 µL5×100 µL-20℃. Avoid freeze/thaw cycle.F665904B10×PCR Buffer 1.8 mL5×1.8 mL-20℃. Avoid freeze/thaw cycle.Notes: 10×PCR Buffer contains 15mM Mg2+. Product IntroductionFastStar DNA Polymerase is a mixture of anti-Taq enzyme monoclonal antibody and CW0680 Taq DNA Polymerase for Hot Start PCR.When using Taq enzyme antibody for PCR amplification, Taq enzyme antibody binds to Taq enzyme to inhibit the DNA polymerase activity prior to high temperature denaturation, and effectively inhibits non-specific annealing of primers and non-specific amplification caused by primer dimerization at low temperatures. It can effectively inhibit the non-specific annealing of primers and non-specific amplification caused by primer dimer under low temperature conditions.Taqase antibody denatures the DNA in the initial DNA denaturation step of the PCR reaction, and DNA polymerase activity is restored to achieve the effect of hot start. No special inactivation of Taq enzyme antibody is required for the use of this product, and it can be used under conventional PCR reaction conditions.FastStar DNA Polymerase with 5′→3′ DNA polymerase activity and 5′→3′ exonuclease activity, no 3′→With 5′ exonuclease activity and an enzyme extension rate of 2kb/min, fragments up to 5kb in length can be amplified. The amplified PCR product has an "A" base at the 3′ end, so it can be directly used for T/A cloning. This product is characterized by fast extension speed and high amplification efficiency, and is mainly suitable for PCR amplification of DNA fragments, DNA sequence determination and other experiments.Active DefinitionUsing activated salmon sperm DNA as template/primer, the amount of enzyme required to dope 10 nmol of deoxyribonucleotide into acid-insoluble material was defined as 1 activity unit (U) at 74°C for 30 min.quality controlAfter several column purifications, its purity was greater than 99% by SDS-PAGE; no exogenous nuclease activity was detected;PCR method detects no host residual DNA; efficiently amplifies single-copy genes in the human genome.UsageThe following example shows the PCR reaction system and reaction conditions for amplifying a 1kb fragment using human genomic DNA as a template, which should be improved and optimized according to the template, primer structure and size of the target fragment in actual operation.Reagent50 µl Reaction systemfinal concentration10× PCR Buffer5 µl1×dNTP Mix，10 mM each1 µl200 µM eachForward Primer，10 µM2 µl0.4 µMReverse Primer，10 µM2 µl0.4 µMTemplate DNA<0.5 µg<0.5 µg/50 µlFastStar DNA Polymerase,5U/µl0.25-0.5 µl1.25-2.5 U/50 µlddH₂Oup to 50 µlNote: The reaction solution can be prepared at room temperature; the reagents must be placed on ice.2.PCR reaction conditionsAttention:1) In general, the annealing temperature in the experiment is 5°C lower than the melting temperature Tm of the amplification primer, and when the ideal amplification efficiency cannot be obtained, the annealing temperature is appropriately lowered.2) The extension time should be set according to the size of the amplified fragment.3) The number of cycles can be set according to the downstream application of the amplification product. If the number of cycles is too low, the amplification is insufficient; if the number of cycles is too high, the chance of mismatch will increase and the non-specific background will be serious. Therefore, the number of cycles should be minimized under the premise of ensuring the product yield... Read More | Product contentF665766Component5 mLStorageF665766A2×Fast Probe Mixture5×1 mL-20℃. Avoid freeze/thaw cycle.F665766BddH2O5×1 mL-20℃. Avoid freeze/thaw cycle.Product IntroductionFast Probe Mixture is a pre-mixed system for real-time fluorescence PCR by probe method (TaqMan, Product contentF665766Component5 mLStorageF665766A2×Fast Probe Mixture5×1 mL-20℃. Avoid freeze/thaw cycle.F665766BddH2O5×1 mL-20℃. Avoid freeze/thaw cycle.Product IntroductionFast Probe Mixture is a pre-mixed system for real-time fluorescence PCR by probe method (TaqMan, Molecular Beacon, etc.), with a concentration of 2×, including Fast Taq DNA Polymerase, PCR Buffer, dNTPs, Mg2+ and so on, which is easy and convenient to operate. It is mainly used for the detection of genomic DNA target sequence and cDNA target sequence after RNA reverse transcription. The Fast Taq DNA Polymerase contained in this product can effectively reduce the non-specific amplification generated by the non-specific binding of primers and templates or primer dimerization at room temperature, and the activation of the enzyme only needs to be incubated at 95 ℃ for 30 s. The whole PCR reaction process can save about 40 minutes compared with the ordinary reaction, which greatly shortens the reaction time of PCR. The combination of unique PCR buffer system and fast hot start enzyme effectively inhibits the generation of non-specific products and significantly improves the PCR amplification efficiency with stronger fluorescence signal, higher sensitivity and wider linear range. The product has a wide range of applications and can be used for both normal and rapid quantitative PCR programs.ROX dye is used to correct the fluorescence signal error generated between wells of a quantitative PCR instrument, and is generally used in Real Time PCR amplifiers from ABI, Stratagene, and other companies. The excitation optics vary from instrument to instrument, so the concentration of ROX dye must be matched to the corresponding fluorescence quantitative PCR instrument.Instruments that do not require ROX calibration (F665766):Roche LightCycler 480, Roche LightCyler 96, Bio-rad iCyler iQ, iQ5, CFX96 and others.Instruments that require Low ROX calibration (F665768):ABI Prism7500/7500 Fast, QuantStudio®3 System, QuantStudio®5 System, QuantStudio®6 Flex System, QuantStudio®7 Flex System, ViiA 7 system. Stratagene Mx3000/Mx3005P, Corbett Rotor Gene 3000, and more.Instruments that require High ROX calibration (F665774):ABI Prism 7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus, and others.matters needing attention1. Before use, please mix gently by turning up and down, avoid foaming as much as possible, and use after brief centrifugation.2. Avoid repeated freezing and thawing of this product, repeated freezing and thawing may degrade the product performance. This product can be stored for long term at -20℃, protected from light. If frequent use is required within a short period of time, it can be stored at 2-8℃.UsageThe following examples are conventional PCR reaction systems and reaction conditions, which should be improved and optimized according to the template, primer structure and target fragment size in actual operation.1.PCR reaction systemReagents50 µl Reaction systemFinal concentration2×Fast Probe Mixture25 µl1×Forward Primer，10 µM1 µl0.2 µM¹⁾Reverse Primer，10 µM1 µl0.2 µM¹⁾Probe，10 µM1 µl0.2 µM²⁾Template DNA2 µl³⁾ 50×Low ROX or High ROX（optional）⁴⁾1 µl1×ddH₂Oup to 50 µlNote: 1) Usually the primer concentration of 0.2µM can get better results, and 0.1-1.0µM can be used as a reference for setting the range. 2) The final concentration of the probe used is related to the fluorescent quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, so please refer to the instruction manual of the instrument or the specific requirements of the use of each fluorescent probe for the adjustment of the concentration in actual use.(3) Usually the amount of DNA template is 10-100ng genomic DNA or 1-10ng cDNA as a reference. Since the templates of different species contain different copy numbers of target genes, the templates can be subjected to gradient dilution to determine the optimal amount of template to be used.(4) The excitation optical system varies from instrument to instrument, choose to add 50×Low ROX or 50×High ROX according to the instrument using fluorescence quantification.2. PCR reaction program:A two-step PCR reaction program is recommended, and this program is set up using the ABI 7500 Fluorescent Quantitative PCR Instrument as a reference.Note: 1) The enzyme used in this product must be pre-denatured at 95°C for 30s to achieve enzyme activation. Under this condition, most of the templates can be well unchained. For templates with high GC content and complex secondary structure, the pre-denaturation time can be extended to 1-4 minutes in order to make the starting template fully unchained.(2) It is recommended to use two-step PCR reaction program, if you can not get good experimental results due to the use of primers with lower Tm values, etc., you can try to carry out a three-step PCR amplification, annealing temperature, please use the range of 56 ℃ - 64 ℃ for as a reference for the setting... Read More | Inquire | Purity>97% SDS-PAGE.FunctionReceptor for interleukin-2 | Purity>95% SDS-PAGE.FunctionProbable cell adhesion protein |