| Description | Product InfoQuality Control Tests:Nuclease contamination: assayed by testing for linear KDNA and linear plasmid DNA formation. Incubation of 1 µg of catenated KDNA or supercoiled pUC19 DNA for 4 hrs. at 37°C (under gyrase assay conditions and with or without ATP).Assay Conditions:Product InfoQuality Control Tests:Nuclease contamination: assayed by testing for linear KDNA and linear plasmid DNA formation. Incubation of 1 µg of catenated KDNA or supercoiled pUC19 DNA for 4 hrs. at 37°C (under gyrase assay conditions and with or without ATP).Assay Conditions:Supercoiling assays are carried out using relaxed pBR-322 DNA under conditions specified in the protocol sheets provided with the product. We also perform assays using kDNA as substrate. For details on this decatenation/supercoiling assay please refer to our technical literature (link provided above). One unit of gyrase activity will supercoil 0.2 ug of plasmid in 30 min at 37°C.Included Materials:A 5X assay uffer and dilution buffer is included with the enzyme. These are also sold separately as 5X Gyrase Assay Buffer Gyrase Dilution Buffer (please enquire).Purification of Gyrase A and B SubunitsDescriptionHighly purified DNA Gyrase is offered as a holoenzyme (contains both A,B subunits). This is an excellent reagent for supercoiling plasmids in vitro or for novel drug screens.Shipping&storageThe enzyme is shipped on dry ice and should be stored at -70°C. We also recommend that the enzyme be aliquoted after the first thaw (repeated rounds of freeze/thaw may cause loss of activity); the enzyme activity is stable for 1-3 days on ice... Read More | Inquire | A general purpose purified albumin, suitable for Westerns, enzyme systems and as a protein supplement | Es Taq DNA Polymerase is an optimized mixed enzyme of Taq and Pfu DNA Polymerase, with 5 '→ 3' DNA polymerase activity, 5 '→ 3' exonuclease activity, and 3 '→ 5' exonuclease activity. Compared with Taq DNA Polymerase, Es Taq DNA Polymerase has excellent performance of high Es Taq DNA Polymerase is an optimized mixed enzyme of Taq and Pfu DNA Polymerase, with 5 '→ 3' DNA polymerase activity, 5 '→ 3' exonuclease activity, and 3 '→ 5' exonuclease activity. Compared with Taq DNA Polymerase, Es Taq DNA Polymerase has excellent performance of high amplification efficiency and low mismatch rate, and can efficiently amplify DNA fragments. Most of the PCR products amplified with this product contain an "A" base at the 3 'end, which can be directly used for T/A cloning. This product is suitable for conventional PCR reactions and gene cloning reactions that require high fidelity. E665597Component500 UStorageE665597AEs Taq DNA Polymerase, 5 U/µL 100 µL -20℃. Avoid freeze/thaw cycle.E665597B10×PCR Buffer 1.8 mL -20℃. Avoid freeze/thaw cycle.Activity definition:Using activated salmon sperm DNA as a template/primer, the amount of enzyme required to incorporate 10 nmol of deoxyribonucleotide into acidic insoluble substances is defined as 1 active unit (U) at 74 ℃ for 30 minutes.Quality control:After multiple column purifications, SDS-PAGE detected a purity of over 99%; No exogenous nuclease activity detected; PCR method for detecting residual DNA without host; Can effectively amplify single copy genes in the human genome; Store at room temperature for one month without significant changes in activity.1. PCR reaction system Reagent 50 µlReaction system Final concentration 10×PCR Buffer 5 µL 1× dNTP Mix,10 mM each 1 µL 200 µM each 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 Es Taq DNA Polymerase,5 U/µl 0.25-0.5 µl 1.25-2.5U/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 ℃ 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 Es 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 | Purity>97% by SDS-PAGE and HPLC analyses.FunctionPlays an important role in the organization of the cytoskeleton (By similarity). Binds to and sequesters actin monomers (G actin) and therefore inhibits actin polymerization. Seraspenide inhibits the entry of hematopoeitic pluripotent stem cells Purity>97% by SDS-PAGE and HPLC analyses.FunctionPlays an important role in the organization of the cytoskeleton (By similarity). Binds to and sequesters actin monomers (G actin) and therefore inhibits actin polymerization. Seraspenide inhibits the entry of hematopoeitic pluripotent stem cells into the S-phase... Read More |