| Description | Product introductionFirefly luciferase is an enzyme that catalyzes production of light from luciferin in the presence of Mg2+-ATP and oxygen. The reaction of this enzyme with luciferin, ATP, and O2 results in the emission of light. Luciferase activity can be inhibited by general anesthetics Product introductionFirefly luciferase is an enzyme that catalyzes production of light from luciferin in the presence of Mg2+-ATP and oxygen. The reaction of this enzyme with luciferin, ATP, and O2 results in the emission of light. Luciferase activity can be inhibited by general anesthetics including isoflurane and ketamine/medetomidine thereby affecting the sensitivity of bioluminescence imaging.ApplicationThe reaction of this enzyme with luciferin, ATP, and O2 results in the emission of light. Luciferase can be used to detect trace amounts of ATP. Firefly luciferase is also one of the most commonly utilized reporter genes for the study of gene expression. The bioluminescent reaction catalyzed by luciferase is one of the most sensitive analytical tools for measuring gene expression. < or equal to one femtomole of ATP can be detected using 0.2 µg of luciferase... Read More | Product Content:F665667Component5 mL40 mLStorageF665667A2×Flash PCR MasterMix (Dye) 5×1 mL 40×1 mL-20℃. Avoid freeze/thaw cycle.F665667BddH2O 5×1 mL40×1 mL-20℃. Avoid freeze/thaw cycle. Products Introduction This product is a premixed system consisting of a new Product Content:F665667Component5 mL40 mLStorageF665667A2×Flash PCR MasterMix (Dye) 5×1 mL 40×1 mL-20℃. Avoid freeze/thaw cycle.F665667BddH2O 5×1 mL40×1 mL-20℃. Avoid freeze/thaw cycle. Products Introduction This product is a premixed system consisting of a new high efficient fast DNA Polymerase, Mg2+, dNTPs, and PCR stabilizers and enhancers at 2× concentration. It is a new rapid DNA polymerase developed by CombiSigma with high amplification speed and stability. The extension speed is up to 5 s/kb, and the PCR can be completed in as little as 15 minutes, while longer fragments (>3 kb) or complex templates can be extended at a speed of 10-30 s/kb or a higher number of cycles. The unique MasterMix formula makes the whole reaction system very stable, while complex templates can be amplified effectively, and more than 98% of PCR amplification can be successful in one run. Simply add the DNA template and primers and top up with water to minimize human error, contamination and time.The dye (blue) has been added to the product and it is ready for electrophoretic detection at the end of the reaction. The PCR product is amplified with an 'A' base at the 3′ end and can therefore be used directly for T/A cloning and is suitable for use in the CombiVerge Seamless Cloning Kit, T4 Ligation Kit and sensory products.This product is mainly suitable for ultra-fast PCR, complex templates, complex secondary structures, gene cloning and large-scale genetic testing that requires high fidelity. quality control No exogenous nuclease activity was detected; no host residual DNA was detected by PCR; single-copy genes in various genomes could be amplified efficiently. UsageThe following is an example of a PCR reaction system and reaction conditions for amplifying a 1 kb fragment using human genomic DNA as a template, which should be improved and optimized according to the template, primer structure and size of the target fragment in actual operation.PCR reaction system Note: 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 concentration can be increased; if a non-specific reaction occurs, the primer concentration can be decreased to optimize the reaction system.PCR reaction conditions Note: 1) Note: For simple templates, the pre-denaturation time can be controlled at 30 s-1 min, for complex templates such as bacterial fluids, the pre-denaturation time can be increased to 2 min.Optimization of parameter settings 1. 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 ng 1. 30-35 number of cycles2. Primer concentration setting: The primer concentration can be set between 0.1 µM and 1.0 µM. A low 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 is 5℃ lower than the melting temperature of amplification primer Tm, so the annealing temperature can be lowered appropriately when the desired amplification efficiency cannot be obtained; the annealing temperature can be raised appropriately when non-specific reaction occurs. For complex templates, it is necessary to adjust the annealing temperature to achieve efficient amplification.4. Extension time setting: The extension time should be set according to the size of the amplified fragments. The following extension times are recommended: simple templates such as plasmids: 5-15 s/kb; regular genomes, cDNA templates: 10-15 s/kb; complex templates, crude templates: 20-30 s/kb; (the extension time should not be too short and should be at least 5 s/kb, but should not exceed 30 s/kb).5. Number of cycles: 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 | The content of this cell is too long for an XLSX file (more than 32767 characters). Please use the CSV format for this export | The Leuconostoc GPDH exhibits dual coenzyme specificity, namely NAD and NADP (Olive and Levy, Biochem., 6, 730 730, 1967). When assayed under conditions that are optimal for the particular coenzyme, the ratio of observed catalytic activity is NAD/NADP = 1.8 | 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 |