| Description | The Anstart Taq-D qPCR Master Mix (2×, with UDG) is a ready-to-use, probe-based qPCR master mix designed for sensitive and contamination-resistant fluorescence quantitative PCR detection in single or multiplex DNA targets. This product provides a simple, rapid, and highly sensitive PCR The Anstart Taq-D qPCR Master Mix (2×, with UDG) is a ready-to-use, probe-based qPCR master mix designed for sensitive and contamination-resistant fluorescence quantitative PCR detection in single or multiplex DNA targets. This product provides a simple, rapid, and highly sensitive PCR detection system, enabling real-time fluorescence PCR amplification in 35–40 minutes.Core ComponentsThe master mix contains:dNTP Mix (including dATP, dTTP, dCTP, dGTP, and dUTP)Antibody-modified hot-start DNA polymerase (Anstart Taq-D)UDG (Uracil-DNA Glycosylase)Users only need to add primers, probes (or fluorescent dyes), and template DNA to perform fluorescence-based qPCR.UDG Contamination Control MechanismDuring the initial 50°C, 2 min incubation, UDG cleaves the N-glycosidic bond between uracil and the sugar-phosphate backbone in any contaminating PCR products containing dUTP, releasing free uracil.Subsequent 95°C, 2.5 min heat treatment inactivates UDG while further degrading the phosphodiester backbone, effectively eliminating carryover contamination from uracil-containing PCR products.UDG acts on both single- and double-stranded DNA but is inactive against RNA.Compatible InstrumentsABI 7500, Qiagen QiaQuant 5-plexFor other instruments not listed, this kit has not been fully validated. If required, please contact our technical support team for further assistance.Protocol1. Reagent Preparation (Reagent Setup Area)1.1 Thaw the master mix at -20 ± 5°C, equilibrate to room temperature, vortex briefly, and centrifuge at low speed for 15 sec before use.Recommendation: Aliquot PCR reagents to avoid repeated freeze-thaw cycles.1.2 Prepare the PCR reaction mix according to the table below:ComponentVolume/25µLVolume/50µL2× Anstart Taq-D qPCR Master Mix12.5 µL25 µLPrimer-probe Mix1 µL2 µLTemplates5 µL10 µLddH2OUp to 25 µLUp to 50 µLTotal25 µL50 µLNote 1: Primer and template volumes can be adjusted as needed.Note 2: For fast cycling programs, a 25 µL reaction volume is recommended.1.3 Mix gently by pipetting or brief vortexing, then centrifuge briefly to collect liquid at the tube bottom.1.4 Transfer PCR tubes to the thermal cycler and initiate the reaction.2. Sample Preparation (Sample Prep Area)Add the appropriate sample volume as required.3. PCR Amplification (Amplification Area)3.1 Preheat the instrument and verify performance.3.2 Load prepared PCR tubes into the instrument and record tube positions. Set fluorescence channels as needed.Recommended Cycling ProgramsStandard ProgramStepCyclesTemp.TimeData Acquisition1150℃2minNo2195℃2min 30sNo34594℃10sNo44555℃40sYes5125℃10sNoFast ProgramStepCyclesTemp.TimeData Acquisition1150℃2minNo2195℃30sNo34594℃5sNo44555℃15sYes5125℃10sNoFluorescence acquisition can be set for 2-step or 3-step PCR depending on experimental needs.Optimal primer concentration: Typically 0.2 µM (Tm ≥ 55°C).Adjust annealing temperature and primer concentration for optimal amplification efficiency... Read More | Bovine pancreatic deoxyribonuclease is an endonuclease which splits phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide yielding polynucleotides with free hydroxyl group at the 3' position and phosphate group at the 5' position. The average chain length of a limit digest is aBovine pancreatic deoxyribonuclease is an endonuclease which splits phosphodiester linkages, preferentially adjacent to a pyrimidine nucleotide yielding polynucleotides with free hydroxyl group at the 3' position and phosphate group at the 5' position. The average chain length of a limit digest is a tetranucleotide.Used for the removal of DNA from protein samples. Deoxyribonuclease I from bovine pancreas has been used in a study to compare several procedures for reducing RNase contamination in preparations of DNase. Deoxyribonuclease I from bovine pancreas has also been used in a study to investigate the effect of the composition of sodium dodecyl sulfate preparations on the renaturation of enzymes after polyacrylamide gel electrophoresis... 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＞97% by SDS-PAGE and HPLC analyses.FunctionPigment epithelium-derived factor (PEDF) is encoded by the SERPINF1 gene in humans and found in verebrates. It is a secreted phosphoglycoprotein that belongs to the clade F subfamily, serpin superfamily of proteinase inhibitors. The PEDF is a Purity＞97% by SDS-PAGE and HPLC analyses.FunctionPigment epithelium-derived factor (PEDF) is encoded by the SERPINF1 gene in humans and found in verebrates. It is a secreted phosphoglycoprotein that belongs to the clade F subfamily, serpin superfamily of proteinase inhibitors. The PEDF is a noninhibitory serpin with neurotrophic, anti-angiogenic, and anti-tumorigenic properties. It is synthesized as a 418 a.a. about 50kDa precursor that contains a 19 a.a. signal sequence and a 399 a.a. mature region that shows a pyroglutamate at Gln20. Like other serpins, it contains three β-sheets, 810 α-helices, and a C-terminal RCL (reactive center loop). Unlike other serpins with Ser protease inhibiting activity. PEDF has functions of inducing extensive neuronal differentiation in retinoblastoma cells, inhibiting of angiogenesis. As it does not undergo the S (stressed) to R (relaxed) conformational transition characteristic of active serpins, it exhibits no serine protease inhibitory activity. PEDF is researched as a therapeutic candidate for treatment of such conditions as choroidal neovascularization, heart disease, and cancer... Read More |