| Description | F665646 Component 5 ml 40 ml Storage F665646A 2×Flash Hot Start MasterMix (Dye) 5×1 mL 40×1 mL -20℃. Avoid freeze/thaw cycle. F665646B ddH₂O 5×1 mL 40×1 mL -20℃. Avoid freeze/thaw cycle.Products IntroductionThis product is a premixed system consisting of a F665646 Component 5 ml 40 ml Storage F665646A 2×Flash Hot Start MasterMix (Dye) 5×1 mL 40×1 mL -20℃. Avoid freeze/thaw cycle. F665646B ddH₂O 5×1 mL 40×1 mL -20℃. Avoid freeze/thaw cycle.Products IntroductionThis product is a premixed system consisting of a new type of highly efficient DNA polymerase, PCR Buffer, Mg2+, dNTPs, and PCR stabilizers and enhancers at a concentration of 2 ×. It contains a new type of high efficient hot starter enzyme, which can effectively inhibit the non-specific annealing of primers and non-specific amplification caused by primer dimer under low temperature. The product has a very high amplification speed and stability, the extension speed can be up to 5-15 sec/kb, suitable for rapid PCR reaction, the original MasterMix formula makes the whole reaction system very stable, more than 98% of the PCR amplification can be successful at once, at the same time, complex templates can also be amplified effectively, and can minimize the human error and contamination. The product has added dye (blue), and can be directly detected by electrophoresis after the reaction. Most of the amplified PCR products have an "A" base at the 3′ end, so they can be used directly for T/A cloning. It is mainly used for rapid PCR reactions and gene cloning where high fidelity is required.Quality controlTested to be free of exogenous nuclease activity; PCR method detects no host residual DNA; can efficiently amplify a wide range of baseSingle-copy genes in the genome.UsageThe following is an example of a PCR reaction system and reaction conditions using human genomic DNA as a template, and the actual operation should be based on the template.Plates, primer structures and target fragment sizes were improved and optimized accordingly.PCR reaction systemNote: Please use the final concentration of 0.1-1.0 µM as a reference for setting the range of primer concentration. If the amplification efficiency is not high, the primer can be increased.concentration; when a non-specific reaction occurs, the primer concentration can be reduced, thus optimizing the reaction system.PCR reaction conditionsNote: If the amplified sample is bacterial liquid, add "pre-denaturation 95℃ for 5min" step.Optimization of parameter settings1. Template DNA amount setting:Excessive amounts of template may result in non-specific amplification or smear. The recommended amount of template DNA in a 50 µl PCR reaction system is as follows:-Human genomic DNA 5 ng-500 ng-Escherichia coli genomic DNA 50 pg-100 ng-plasmid DNA 10 pg-1 ng2. Primer concentration setting:The primer concentration can be set between 0.1 µM and 1.0 µM. Too low a primer concentration may result in low amplification products. Too high a primer concentration will inhibit specific amplification and may result in non-specific amplification.3. Annealing temperature setting: In general, the annealing temperature in the experiment is 5℃ lower than the melting temperature of the amplification primer, Tm, which is not able to get the ideal amplification efficiency.The annealing temperature can be lowered appropriately in the case of non-specific reactions; the annealing temperature can be increased appropriately in the case of non-specific reactions. For complex templates, the annealing temperature needs to be adjusted to achieve efficient amplification.4. Extension time setting: The extension time should be set according to the size of the amplified fragments. The following are recommended extension times:Simple templates such as plasmids: 5-15 s/kb; Conventional genome, cDNA templates: 10-15 s/kb; Complex templates, crude extraction templates: 20-30 s/kb;(The extension time should not be too short it should be at least 5 s/kb and not more than 30 s/kb).5. Cycle number setting: The number of cycles can be set according to the downstream application of the amplified product. If the number of cycles is too low, the amount of amplification will be 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 | 1、Product attributeShelf life: 24 monthsReaction time：long (up to 45 minutes) at 20-37°CLot-to-lot variation：<10%Boiling point : 100℃pH-Value (at 20 °C): 9.0-9.8 Density (20℃) : 1.0302 g/cm³Water solubility: easily solubleAppearance: colourless to 1、Product attributeShelf life: 24 monthsReaction time：long (up to 45 minutes) at 20-37°CLot-to-lot variation：<10%Boiling point : 100℃pH-Value (at 20 °C): 9.0-9.8 Density (20℃) : 1.0302 g/cm³Water solubility: easily solubleAppearance: colourless to light yellow liquidOdour: odourlessIncubation temperature: 20-37 °CLight sensitiveHeat sensitive 2、Requirements for storage rooms and vessels1.Keep container tightly closed.2.Keep cool. protected from light3. Do not store together with: Oxidizing agent. 4. Contaminated or leaked out substrate solution from damaged bottles should not be used anymore and has to be destroyed.5. Use isolated containers with some cool bags for transport.6. Spontaneous decay will increase the background. If stored at room temperature, the velocity of the decay will increase. Thus, both storage and transport at room temperature should be avoided. Nevertheless, the activity of the solution is not affected by storage at room temperature. The solution still works beyond the expiry date, but some applications, especially those including visual evaluation, may be hampered by increased background. 3、Effective Components and Principle of FunctionIn different buffer solutions (pH = 9.5), with supplementation if required, the effective componentpara nitrophenyl phosphate (pNPP) is dissolved. Alkaline Phosphatase transfers the phosphate residue to an acceptor. Under alkaline conditions a yellow colour occurs, resulting from the formed nitrophenol. 4、Biosafety informationThis mixture is not classified as hazardous in accordance with Regulation (EC) No 1272/2008； 5、Advantage1. Signal yield comparable to competitor ready-to-use solutions2. Broad measurement range3. Very low background signals4. Very low blank drift during long-term storage (<0.15 AU within 24 months)5. High colour stability after reaction stop with this product and other commonly used stopping solutions 6、Instruction for usageFor bottling consider the following instructions:• Work in a dust free and darkened room.• Keep the solution as cool as possible.• Avoid contact of the solutions with any metal parts• Clean all instruments and vessels very extensively.• Wear powder-free gloves during bottling.• Close the bottles immediately to minimize the influence of light and dust.• Use clean bottles that are impermeable to light made from HDPE or PP. 7、 General Instructions for the Use in Blotting Systems Only qualified laboratory staff, who are familiar with the basics of immunological methods, are allowed to use these solutions.The substrate solutions can be used in qualitative and quantitative ELISA procedures.When using 96-well microtiter plates, adding 100 µL of substrate per well after incubation and washing is recommended. After substrate incubation the reaction can be stopped and the photometric measurement can be carried out. Using higher incubation temperatures (37° C) may shorten the incubation time. The reaction can be stopped by using the special developed solution stop. The use of other commercially available stop solutions cannot safely exclude a further increase of the signal. Addition of a stopping solution does not change the general shape of the spectrum. The unstopped and the stopped solution should be measured at 405 nm and the background correction: should be measured at 620 nm... Read More | Inquire | Purity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：Involved in the high-affinity maltose membrane transport system MalEFGK. Initial receptor for the active transport of and chemotaxis toward maltooligosaccharides.Epitope tagging offers an easy and universalPurity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：Involved in the high-affinity maltose membrane transport system MalEFGK. Initial receptor for the active transport of and chemotaxis toward maltooligosaccharides.Epitope tagging offers an easy and universal strategy for the identification and purification of proteins derived by recombinant DNA technology. The insertion of a Maltose Binding Protein (MBP) tag creates a stable fusion product that does not interfere with the bioactivity of the protein or with the biodistribution of the MBP tagged product... Read More | Purity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：CNN1 is a member of the calponin family. CNN1 is a thin filament-associated protein which is involved in the regulation and modulation of smooth muscle contraction. CNN1 is able to bind to actin, calmodulinPurity: >90%, by SDS-PAGE visualized with Coomassie® Blue Staining.Description：CNN1 is a member of the calponin family. CNN1 is a thin filament-associated protein which is involved in the regulation and modulation of smooth muscle contraction. CNN1 is able to bind to actin, calmodulin, troponin C and tropomyosin. Prevention of actomyosin Mg-ATPase activity is a result of interaction between calponin and actin... Read More |