| Description | Product contentG665762Component5 mLStorageG665762A2×GoldStar Probe Mixture5×1 mL-20℃. Avoid freeze/thaw cycle.G665762B50×High ROX200 µL-20℃. Avoid freeze/thaw cycle.G665762CddH2O5×1 mL-20℃. Avoid freeze/thaw cycle.Product IntroductionGoldStar Probe Mixture Product contentG665762Component5 mLStorageG665762A2×GoldStar Probe Mixture5×1 mL-20℃. Avoid freeze/thaw cycle.G665762B50×High ROX200 µL-20℃. Avoid freeze/thaw cycle.G665762CddH2O5×1 mL-20℃. Avoid freeze/thaw cycle.Product IntroductionGoldStar Probe Mixture is a premixed system for real-time fluorescence quantitative PCR by probe method (TaqMan, Molecular Beacon, etc.), with a concentration of 2×, containing GoldStar Taq DNA Polymerase, PCR Buffer, dNTPs and Mg2+, which is easy and convenient to operate. It is mainly used for genomic DNA target sequence and RNA reverse transcription post-cDNA target sequence detection, such as gene expression analysis, copy number analysis, SNP genotype analysis, etc. It is suitable for fluorescence quantification by different types of probe methods. GoldStar Taq DNA Polymerase is a chemically modified, new and highly efficient hot-start enzyme, which has no polymerase activity at room temperature and effectively avoids non-specific amplification caused by non-specific binding of primers and templates or primer dimerization at room temperature, and the activation of the enzyme must be incubated at 95℃ for 10 minutes. The unique combination of PCR buffer system and hot-start enzyme significantly improves the amplification efficiency of PCR with stronger fluorescent signal and higher sensitivity to detect single-copy templates. A wider linear range and more accurate quantification of target genes can be obtained with this product. It is suitable for all fluorescent quantitative PCR instruments that do not require ROX as a calibration dye.ROX dye is used to correct the fluorescence signal error generated between wells of a quantitative PCR instrument, and is generally used in Real Time PCR amplifiers from ABI, Stratagene, and other companies. The excitation optics vary from instrument to instrument, so the concentration of ROX dye must be matched to the corresponding fluorescence quantitative PCR instrument.Instruments that do not require ROX calibration(G665762):Roche LightCycler 480, Roche LightCyler 96, Bio-rad iCyler iQ, iQ5, CFX96 and others.Instruments requiring Low ROX calibration(G665762):ABI Prism7500/7500 Fast, QuantStudio®3 System, QuantStudio®5 System, QuantStudio®6 Flex System, QuantStudio®7 Flex System, ViiA 7 system. Stratagene Mx3000/Mx3005P, Corbett Rotor Gene 3000, and more.Instruments requiring High ROX calibration(G665762):ABI Prism 7000/7300/7700/7900, Eppendorf, ABI Step One/Step One Plus, and others.matters needing attention1. Before use, please mix it gently by turning it up and down, avoid foaming as much as possible, and use it after centrifugation for a short time.2. Avoid repeated freezing and thawing of this product, repeated freezing and thawing may degrade the product performance. This product can be stored for a long time at -20℃, protected from light. If frequent use is required in the short term, it can be stored at 2-8℃.UsageThe following examples are conventional PCR reaction systems and reaction conditions, which should be improved and optimized according to the template, primer structure and target fragment size in actual operation.1.PCR reaction systemReagents50 µl Reaction systemFinal concentration2×GoldStar Probe Mixture25 µl1×Forward Primer，10 µM1 µl0.2 µM ¹⁾Reverse Primer，10 µM1 µl0.2 µM ¹⁾Probe，10 µM1 µl0.2 µM ²⁾Template DNA2 µl³⁾ 50×Low ROX or High ROX（optional）⁴⁾1 µl1 ×ddH₂Oup to 50 µlNote: 1) Usually, the primer concentration of 0.2µM can get better results, and 0.1-1.0µM can be used as a reference for setting the range. 2) The concentration of the probe used is related to the fluorescent quantitative PCR instrument used, the type of probe, and the type of fluorescent labeling substance, so please refer to the instruction manual of the instrument or the specific requirements of the use of each fluorescent probe for the adjustment of the concentration during the actual use.(3) Usually the amount of DNA template is 10-100ng genomic DNA or 1-10ng cDNA as a reference. Since the templates of different species contain different copy numbers of target genes, the templates can be subjected to gradient dilution to determine the optimal amount of template to be used.(4) The excitation optical system varies from instrument to instrument, choose to add 50× Low ROX or 50× High ROX according to the instrument using fluorescence quantification.2.PCR reaction programCaution! The pre-denaturation reaction of this product must be completed at 95°C for 10 minutes!Two-step PCR::Note: 1) The hot-start enzyme used in this product shall be activated by the enzyme under the condition of pre-denaturation 95℃ and 10min.(2) It is recommended to use two-step PCR reaction program, if you do not get good experimental results due to the use of primers with lower Tm values, etc., you can try to carry out three-step PCR amplification, and the annealing temperature, please use the range of 56 ℃ - 64 ℃ as a setting reference... Read More | Inquire | Store at +4°C. Store under desiccating conditions. The product can be stored for up to 12 months | 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 | Ribonuclease T1 is an endoribonuclease, highly specific for the cleavage of RNA or deaminated RNA between guanosine 3'-phosphate residues (or inosine 3'-phosphate) and the 5'-OH residues of adjacent nucleotides with the formation of the corresponding intermediate 2', 3'-cyclic phosphates. It cleavesRibonuclease T1 is an endoribonuclease, highly specific for the cleavage of RNA or deaminated RNA between guanosine 3'-phosphate residues (or inosine 3'-phosphate) and the 5'-OH residues of adjacent nucleotides with the formation of the corresponding intermediate 2', 3'-cyclic phosphates. It cleaves single-stranded RNA releasing oligonucleotides from the guanosine 3'-phosphate termini. The enzyme has a molecular weight of 11 kDa. The optimum pH is 7.5. RNase T1 is inhibited by Ag+, Zn2+, Cu2+, and Hg2+ at 1 X 10-3 M. The stimulatory effects of both histidine and EDTA are attributed to chelation of contaminating inhibitor cations. The enzyme assay is essentially the method of Egami et al., Prog. in Nucleic Acid Res. and Molec. Biol., III, 59 (1964) based upon the release of acid soluble oligonucleotides following the digestion of yeast RNA.Ribonuclease T1 (RNase T1) from Aspergillus oryzae is used to digest denatured RNA prior to sequencing and is used for protein folding studies. ApplicationRibonuclease T1 has extensive applications in molecular cloning and DNA sequencing. Because of its specificity it has been a commonly used cleavage enzyme for the determination of structure, nearest neighbor frequencies, and RNA sequencing. The enzyme has further application in the preparation of nucleoside 2',3'-cyclic phosphates, the synthesis of oligonucleotides, and the removal of RNA from DNA preparations. The enzyme is also used as a non-mammalian source of RNase in various applications... Read More |