Takara Bio's R&D scientists are expert enzymologists who are continually working to improve enzymes and buffers for PCR, real-time PCR, and RT-PCR. As a result, Takara Bio's enzymes are designed to make PCR work for you.
You expect a lot from your real-time PCR reagents- they need to perform at a high level and provide reproducibility, instrument compatibility, and sensitivity at a great value. Takara's real-time PCR master mixes are designed with these issues in mind. Whether your real-time experiments are routine or if you're having difficulties, Takara has a real-time PCR kit for you.
Use the tabs below to explore all of the features of Takara's real-time PCR kits.
High assay efficiency is critical to obtaining accurate qPCR data. Several factors can affect the overall efficiency of your qPCR including the kit used for amplification. Takara's real-time PCR master mixes provide excellent and consistent efficiency compared to other commercially available qPCR kits.
Figure 1. PCR efficiency was assessed using SYBR®Premix Ex Taq™ (Tli RNase H Plus) and other commercially available SYBR® master mixes (Company A, B, and C). Three portions of the ACTB gene were amplified (red: 186 bp; black: 381 bp; blue: 533 bp) from cDNA synthesized using 10 pg to 1 µg of human testes total RNA, and the standard curves were plotted.
Specificity is important- you don't want to detect products that aren't really there, and you don't want your results to be artificially inflated by non-specific amplification products. Good primer design can significantly reduce this problem, but the formulation of your master mix is also important. The enzyme and buffer in each of Takara's real-time PCR master mixes have been optimized to maximize specificity even for difficult amplifications (GC-rich products).
SYBR® Premix Ex Taq™ II provides excellent specificity
Figure 1. Assay specificity was assessed using SYBR® Premix Ex Taq™ II (Tli RNaseH Plus) and other commercially available SYBR® master mixes (Company A and B). cDNA was synthesized from 10 pg to 1 µg of human testes total RNA. The expression of ACTB (186 bp and 381 bp fragments) was measured by real-time PCR using a StepOnePlus™ system (ABI). Non-specific amplification was not observed for reactions with Takara's premix.
SYBR® Premix Ex Taq™ II provides excellent specificity for GC-rich targets
Figure 2. Assay specificity for GC-rich targets was assessed using SYBR® Premix Ex Taq™ II (Tli RNaseH Plus) and other commercially available SYBR® master mixes (Company A and B). cDNA was synthesized from 10 pg to 1 µg of human testes total RNA using PrimeScript Reverse Transcriptase. The expression of C16orf84 (73% GC content) and PLSCR3 (67% GC content) was measured by real-time PCR using a StepOnePlus™ system (ABI). The standard curves indicate that Takara's SYBR master mix provided high efficiency amplification. In addition, non-specific amplification was not observed for reactions with Takara Bio's premix, indicating high assay specificity for these GC-rich targets.
What instrument do you use? Takara Bio's real-time PCR master mixes have been tested for compatibility with a variety of instruments. In addition, the user manuals for qPCR premixes include protocols for commonly used real-time PCR instruments. In addition to the instruments listed below, Takara Bio field testers have successfully used SYBR®Premix Ex Taq™ (Cat. # RR420) on the ABI 7900HT Fast Real-Time PCR System and SYBR®Premix ExTaq™ II (Cat. # RR820) on the BioRad CFX Connect™ Real-Time PCR Detection System.
ABI PRISM® 7000/7700
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR Systems
StepOnePlus™ Real-Time PCR System (Life Technologies)
Smart Cycler® System/Smart Cycler® II System (Cepheid)
Whether you need a high throughput workflow or just want your results fast, Takara Bio's real-time PCR master mixes can reduce your run times. For maximum reaction speed, the master mixes can be used with fast mode thermal cycling conditions.
Premix Ex Taq™ (Probe qPCR) has the same reaction efficiency with extension times of 20, 25, and 30 seconds on an Applied Biosystems 7500 Fast Real-Time PCR System.
Premix Ex Taq™ (Probe qPCR) has the same reaction efficiency with extension times of 20 and 30 seconds on a Roche LightCycler® 480 System.
Takara's real-time PCR master mixes stretch your research dollars without sacrificing quality. Make the switch and you can save hundreds or even thousands of dollars annually. Compare the qPCR reagent you are currently using to the list prices below.
Conversion of mRNA to cDNA can introduce variability-reverse transcriptases have different efficiencies that can be target-dependent. RNA secondary and tertiary structures, variation in priming efficiency, and properties of the reverse transcriptase itself can affect cDNA synthesis. Takara Bio's PrimeScript Reverse Transcriptase is a robust enzyme that can quickly (~ 15 minute reaction time) synthesize high yields of cDNA even from the most difficult RNAs. The PrimeScript RTase kits below generate cDNA that can be used directly in real-time PCR.
If you want to...
Make cDNA to be used in real time PCR using a master mix that includes RTase and primers
Do you want a more streamlined workflow? In one-step real-time RT-PCR, all reactions (cDNA synthesis, PCR amplification, and detection) are performed in a single tube. A one-step protocol is simple and convenient, and minimizes the possibility of reaction contamination. Takara Bio provides one-step real-time RT-PCR kits for both SYBR® Green and TaqMan® probe detection. These kits include PrimeScript™ RTase, an RNase H Minus MMLV-based reverse transcriptase, TaKaRa Ex Taq® HS, a high efficiency hot start PCR enzyme, and a buffer system optimized for one-step RT-PCR.
Efficient detection of human HPRT1 expression using one-step real-time RT-PCR
Figure 1. The One Step SYBR® PrimeScript™ RT-PCR Kit II (Cat. # RR086A) was used to measure the level of HPRT1 and generate a standard curve. Human liver total RNA (6.4 pg - 100 ng) was used as a template and reactions were run following the recommended protocol. Amplification curves were generated for all of the template concentrations. The overlapping melt curves indicate that the same amplification products were generated even when different amounts of template were used. The data generated a linear standard curve within the wide range of RNA template concentrations.