| Description | This product is a reagent kit for reverse transcription of genomic DNA, which can remove genomic DNA in 2 minutes at 42 ℃. Meanwhile, the reverse transcription reagent contains components that inhibit gDNA Remover, and the samples treated with gDNA Remover can be directly subjected to reverse This product is a reagent kit for reverse transcription of genomic DNA, which can remove genomic DNA in 2 minutes at 42 ℃. Meanwhile, the reverse transcription reagent contains components that inhibit gDNA Remover, and the samples treated with gDNA Remover can be directly subjected to reverse transcription reaction to synthesize cDNA.This reagent kit is equipped with a novel and efficient reverse transcriptase HiFiScript, which significantly enhances the transcriptional activity of the enzyme through novel mutation sites. Meanwhile, reverse transcription reaction only takes 15 minutes to complete the synthesis of the first strand of cDNA. 5 × HifiScript RT MasterMix is a reverse transcription premix that contains all the reagents required for reverse transcription, making it easy and fast to operate.H665909Component100 TStorageH665909A10×gDNA Remover Mix 100 µL-20℃. Avoid freeze/thaw cycle.H665909B5×HiFiScript RT MasterMix 400 µL-20℃. Avoid freeze/thaw cycle.H665909CRNase-Free Water 1.5 mL-20℃. Avoid freeze/thaw cycle. Product features1. Quick genome removal: gDNA Remover containing genomic DNA can remove genomic DNA in just 2 minutes.2. Rapid reverse transcription: cDNA first strand synthesis can be completed in 15 minutes.3. Convenient and fast: Ready to use reverse transcription Mix, easy to operate.4. High sensitivity: cDNA first strand can be synthesized using pg level total RNA or mRNA templates.5. Efficient reverse transcription efficiency: Novel mutation sites significantly enhance enzyme activity performance, resulting in higher yields of cDNA.Matters needing attention1. During the operation, RNase contamination should be avoided to prevent RNA degradation or cross contamination during experiments. It is recommended that operators wear masks and disposable gloves, frequently change gloves, and use specialized instruments and consumables.2. Disposable plastic containers should be used as much as possible for experiments. If glassware is used, a 0.1% DEPC (diethyl carbonate 1. ester) aqueous solution should be treated at 37 ℃ for 12 hours, and then sterilized under high pressure at 120 ℃ for 30 minutes before use. Alternatively, glassware should be sterilized under dry heat at 180 ℃ for 60 minutes before use. The sterile water used in the experiment should be treated with 0.1% DEPC and then subjected to high-pressure sterilization.3. The reverse transcription system is prepared on ice for operation to prevent RNA degradation. The enzyme in the reagent kit should be stored at -20 º C as soon as possible after use, and repeated freeze-thaw should be avoided as much as possible.UsageThaw the template RNA on ice; After thawing at room temperature, immediately place the components of the reagent kit on ice. Before use, mix each solution by vortex oscillation and centrifuge briefly before use.1、 Genomic DNA removal reaction1. Prepare a reaction system on ice according to the following table, with a total volume of 10 µ L. To ensure the accuracy of the preparation of the reaction solution, a pre mixed system is first prepared in the amount of reaction number+2, then divided into each reaction tube, and finally RNA samples are added. Reagent 10 µl Reaction system 10×gDNA Remover Mix 1µl RNA Template¹ 10 pg-1 µg RNase-Free Water up to 10 µl Note: 1) If the total RNA amount is greater than 1 µ g, please expand the reaction system proportionally.2. Vortex shake and mix well, briefly centrifuge to collect the solution on the pipe wall to the bottom of the pipe. 3. Incubate at 42 ℃ for 2 minutes (during room temperature reaction, it can be extended to 30 minutes).4. After the reaction is complete, centrifuge briefly and cool on ice.2、 Reverse transcription reaction1. Prepare the reaction system on ice according to the following table, and prepare the reaction solution on ice. To ensure the accuracy of the reaction solution configuration, first prepare a premixed solution in the amount of reaction number+2, and then divide it into 10 batches µ Take 10% of the prepared premix from each reaction tube µ Add to the reaction tube of step 1 that has completed genome removal. Reagent 20 µl Reaction system Step 1 Reaction solution 10µl 5×HiFiScript RTMaster Mix 40µl RNase-Free Water 60µl 2. Mix well and centrifuge briefly to collect the solution on the tube wall to the bottom of the tube.3. cDNA synthesis reaction conditions: Incubate at 37 ℃ for 15 minutes and 85 ℃ for 5 seconds.4. After the reaction is completed, centrifuge briefly and place it on ice before proceeding with the subsequent reaction. If it needs to be stored for a long time, please place it at -20 ℃... 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 | Protein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 10.16 at 1.0 mg/ml for pure C3Molecular Weight187,000 Da (2 chains)General DescriptionRat C3 is purified from pooled normal rat serum. C3 is central to the activation of all three pathways of complement activation (Law, S.K.A. and Reid, KProtein Purity≥85% by SDS PAGEExtinction CoeffA280 nm = 10.16 at 1.0 mg/ml for pure C3Molecular Weight187,000 Da (2 chains)General DescriptionRat C3 is purified from pooled normal rat serum. C3 is central to the activation of all three pathways of complement activation (Law, S.K.A. and Reid, K.B.M. (1995)). Initiation of each pathway generates proteolytic enzyme complexes (C3 convertases) which are bound to the target surface. These enzymes cleave a peptide bond in C3 releasing the anaphylatoxin C3a and activating C3b. For a brief time (~60 µs) this nascent C3b is capable of reacting with and covalently coupling to hydroxyl groups on the target surface. Carbohydrates are the favored target, but protein hydroxyls and amino groups also react. This process of tagging the target surface with C3b is called opsonization. The reactive site in nascent C3b is a thioester (Tack B.J., et al. (1980); Pangburn M.K. and MüllerEberhard H.J. (1980)) and C3b is linked to the target through a covalent ester bond (an amide bond is formed if C3b is attached to amino groups). Most of the C3 activated during complement activation never attaches to the surface because its thioester reacts with water forming fluid phase C3b which is rapidly inactivated by factors H and I forming iC3b. Surface-bound C3b is necessary in all three pathways for efficient activation of C5 and formation of C5b-9 complexes that lyse the target cell membrane. Surface-bound C3b and its breakdown products iC3b and C3d are recognized by numerous receptors on lymphoid and phagocytic cells which use the C3b ligand to stimulate antigen presentation to cells of the adaptive immune system. The end result is an expansion of target-specific B-cell and T-cell populations.Physical Characteristics & StructureThe calculated molecular weight of rat C3 based on its amino acid sequence is 184,111daltons (without the signal peptide) and is similar to that of human C3 (185,000 daltons).The molecular weight of rat C3 as determined by SDS/polyacrylamide gel electrophoresis has been reported by Daha, M.R. et al., (1979) to be 187,000 daltons composed of two disulfide linked chains, alpha chain (123,000 daltons) and beta chain (76,000 daltons). The extinction coefficient of rat C3 (E1%/280nm = 10.16) is calculated based on its amino acid sequence using ProtParam and assumes all pairs of Cys residues form cystines (i.e. a pair of cysteine molecules are joined by a disulfide bond). The theoretical pI of rat C3 is 6.12. The normal plasma concentration of C3 inWistar rats has been reported to be 0.581mg/ml (Daha, M.R. et al., (1979)).FunctionThe biological functions of C3 are described above in the General Description section.GeneticsRat C3 chromosome location 9. The NCBI Gene ID number for rat C3 is 24232 and UniProt accession number is P01026.Precautions/Toxicity/HazardsThis protein is purified from animal plasma/serum and therefore precautions appropriate for handling any animal blood-derived product must be used.ReferencesLaw, S.K.A. and Reid, K.B.M. (1995) Complement 2nd Edition (ISBN 0199633568) Oxford University Press, Oxford.Tack BF, Harrison RA, Janatova J, Thomas ML, Prahl JW. (1980) Evidence for presence of an internal thiolester bond in third component of human complement. Proc Natl Acad Sci U S A. 77:5764-8.Pangburn M.K. and Müller-Eberhard H.J. (1980) Relation of putative thioester bond in C3 to activation of the alternative pathway and the binding of C3b to biological targets of complement. J Exp Med. 152:1102-14.Daha MR, Stuffers-Heiman M, Kijlstra A and Van ES LA. (1979) Isolation and characterization of the third component of rat complement. Immunology 36:63-70... Read More | Product Application：KNK437 has been used: as a heat shock factor 1 (HSF1) inhibitor to study its effects on the inhibition of viability and apoptosis activation in chemoresistant mice cells as an HSF1 inhibitor to study its effects on viability and apoptosis of colorectal cancer cells as a Product Application：KNK437 has been used: as a heat shock factor 1 (HSF1) inhibitor to study its effects on the inhibition of viability and apoptosis activation in chemoresistant mice cells as an HSF1 inhibitor to study its effects on viability and apoptosis of colorectal cancer cells as a heat shock protein 70 (HSP70) inhibitor to study its effects on glutamine-induced HSP70 and inflammatory mediator release... Read More | Inquire |