| Description | Products contentS665634Component1 mL5 mLStorageS665634A2×Super Pfx Master Mix1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle.S665634BddH₂O1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle. Products IntroductionIt is an optimized 2× premixed reagent consisting of Super Pfx DNA Products contentS665634Component1 mL5 mLStorageS665634A2×Super Pfx Master Mix1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle.S665634BddH₂O1 mL 5×1 mL-20℃. Avoid freeze/thaw cycle. Products IntroductionIt is an optimized 2× premixed reagent consisting of Super Pfx DNA Polymerase, Mg2+, dNTPs, and reaction buffer, and requires only the addition of primers and DNA templates for the PCR reaction.Super Pfx DNA Polymerase has a 3´→5´ exonuclease activity, and is approximately 100 times more fidelity than Taq DNA Polymerase. Super Pfx DNA Polymerase has 3´→5´ exonuclease activity and is about 100 times more fidelity than Taq DNA Polymerase, making it ideal for cloning. With the introduction of elongation enhancement technology, this enzyme enables high-speed PCR with an elongation speed of 10 sec/kb and an amplification length of up to 20 kb, while maintaining high fidelity. In addition, this product combines a high success rate with the ability to amplify long fragments that are difficult to amplify, GC-rich regions, and templates that are too large to be amplified. The amplified PCR product does not have an "A" base at the 3´ end and can be directly cloned into a flat-end vector. This product is divided into the standard 2×Master Mix without Loading Dye and the 2×Master Mix with Loading Dye.Product Characteristics1. High-speed PCR possibleThe extension time can be set to 10sec for amplification of target fragments of less than 1kb, and 20-30sec/kb for amplification of target fragments of 1-10kb, which significantly reduces the PCR reaction time compared to conventional reagents.2. High fidelityThe fidelity of Taq DNA polymerase is about 100 times higher than that of Taq DNA polymerase, and long target fragments can be amplified quickly and with high fidelity, and the amplified products can be used for a variety of purposes.3. simple and convenientThis reagent contains all PCR components except primers and templates, which facilitates the operation and improves the reproducibility of the results. In addition, 2×Super Pfx Master Mix (Dye) contains Loading Dye, which can be used for gel electrophoresis directly after the reaction.caveatPrimers and templates with uracil are not suitable for this product.Instructions for usePCR Reactors1. Before preparing the reaction solution, please melt and mix the reagents completely before use.After adding the components, please mix them well and transfer them to the PCR instrument quickly.・The recommended final primer concentration is 0.4-0.5 µM, but for amplification of long fragments of 10 kb or more, a final primer concentration of 0.3 µM can increase the amount of amplified product.PCR reaction program1. Before preparing the reaction solution, please melt and mix the reagents completely before use.take note of 1）Pre-denaturation: for most purified templates, 98℃, 30sec is sufficient; for complex templates, the pre-denaturation time can be extended, not more than 3min. 2）Annealing: In general, the annealing temperature is 3-5°C lower than the primer Tm. When a non-specific reaction occurs, increase the annealing temperature appropriately. If necessary, a temperature gradient can be established to find the optimal temperature for primer-template binding. For high Tm primers, a two-step cycle can be used, combining annealing and extension into one step.3）Extension: for complex genomic samples, extension times are typically 20-30sec/kb, but can be reduced to 10sec/kb for simple templates (plasmids, E. coli, etc.) or complex templates <1kb. cDNAs, or complex templates >6kb, can be increased to 40-50sec/kb, if desired... 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 | Unit Definition One unit will cause a change in A600 of 0.330 per minute at pH 5.7 at 37°C in a 2.0 ml reaction mixture (45 minute assay) | Inquire | Tyrosine decarboxylase catalyzes the removal of the carboxyl group from tyrosine to produce tyramine and carbon dioxide. Pyridoxal 5'-phosphate is a necessary cofactor. By using the apoenzyme prepared from cells grown on a vitamin B6 deficient medium pyridoxal phosphate may be determined. The Tyrosine decarboxylase catalyzes the removal of the carboxyl group from tyrosine to produce tyramine and carbon dioxide. Pyridoxal 5'-phosphate is a necessary cofactor. By using the apoenzyme prepared from cells grown on a vitamin B6 deficient medium pyridoxal phosphate may be determined. The HOLOenzyme may be used to determine tyrosine, phenylalanine and dihydroxyphenylalanine either manometrically or colorimetrically.L-Tyrosine decarboxylase apoenzyme from Streptococcus faecalis has been used in a study to purify and characterize tyrosine decarboxylase and aromatic-L-amino-acid decarboxylase.L-Tyrosine decarboxylase apoenzyme from Streptococcus faecalis has also been used in a study to investigate the stereospecificity of sodium borohydride reduction of tyrosine decarboxylase... Read More |