| 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 | Inquire | Taq MasterMix is a premixed system composed of Taq DNA Polymerase, Mg2+, dNTPs, as well as PCR stabilizers and enhancers, with a concentration of 2 ×. The pre prepared PCR mixture makes the operation simpler and faster, and can minimize human error and contamination to the greatest extent Taq MasterMix is a premixed system composed of Taq DNA Polymerase, Mg2+, dNTPs, as well as PCR stabilizers and enhancers, with a concentration of 2 ×. The pre prepared PCR mixture makes the operation simpler and faster, and can minimize human error and contamination to the greatest extent possible. The original MasterMix formula results in high yield, strong repeatability, and good stability of amplified products. This product has been added with a dye (blue), and can be directly subjected to electrophoresis detection after the reaction is completed. The amplified PCR product has an "A" base attached to its 3 'end, making it suitable for direct use in T/A cloning. Mainly suitable for PCR amplification of DNA, DNA sequencing and other experiments. T665590Component5 mL25 mLStorageT665590A2×Taq MasterMix (Dye)5×1 mL5×5 mL-20℃. Avoid freeze/thaw cycle.T665590BddH₂O5×1 mL5×5 mL-20℃. Avoid freeze/thaw cycle.2×Taq MasterMix contains Taq DNA Polymerase, 3 mM Mg Cl₂ and 400 µM each dNTP. Quality control:After testing, there was no exogenous nuclease activity; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes from multiple genomes.Usage:The following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template. In practical operation, corresponding improvements and optimizations should be made based on the template, primer structure, and target fragment size.1. PCR reaction system Reagent 50 µlReaction system Final concentration 2×Taq MasterMix（Dye） 25 µL 1× Forward Primer，10 µM 2 µL 0.4 µM Reverse Primer，10 µM 2 µl 0.4 µM Template DNA <0.5 µg <0.5 µg/50 µL ddH2O up to 50 µL /Attention: The primer concentration should be between 0.1 and 1.0 as the final concentration µ M serves as a reference for setting the range. In the case of low amplification efficiency, the concentration of primers can be increased; When non-specific reactions occur, the primer concentration can be reduced to optimize the reaction system.2. PCR reaction conditions Step Temperature Time / Pre denaturation 94℃ 2 min / Denaturation 94℃ 30 s 25-35 cycles Anneal 55-65℃ 30 s 25-35 cycles Extend 72℃ 30 s 25-35 cycles Finally extended 72℃ 2 min / Attention:1) In general experiments, if the annealing temperature is 5 ° C lower than the melting temperature Tm of the amplification primer, and the ideal amplification efficiency cannot be achieved, the annealing temperature should be appropriately reduced; When non-specific reactions occur, increase the annealing temperature to optimize the reaction conditions.2) The extension time should be set according to the size of the amplified fragment. The amplification efficiency of Taq DNA Polymerase in this product is 2 kb/min.3) The number of cycles can be set based on the downstream application of the amplification product. If the number of cycles is too small, the amplification amount is insufficient; If there are too many cycles, the probability of mismatches will increase, and non-specific backgrounds will be severe. So, while ensuring product yield, the number of cycles should be minimized as much as possible... Read More | Store at +4°C. Store under desiccating conditions. The product can be stored for up to 12 months | Inquire |