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DTI FabGreen HiFid Taq HS II Premix is a 2X concentration reagent optimized for intercalator-based real-time PCR. It includes green intercalator dye at a concentration appropriate for real time monitoring for easy reaction setup and Tli RNase H to reduce PCR inhibition from residual mRNA in cDNA templates. The modified buffer and hot start PCR enzyme with an anti-Taq antibody ensure high specificity and reliable real-time PCR results.

Specifications

Documents

Applications

Brand	Catalogue Number	Pack Size
DTI Green HiFid Taq HS II Premix	DT0601.100	100 rxn (20 μl volume)
DTI Green HiFid Taq HS II Premix	DT0601.500	500 rxn (20 μl volume)

• High specificity minimizes primer-dimer formation and nonspecific amplification

• Excels with GC-rich targets

• 2X master mix provides convenience

• High sensitivity: detects fewer than 100 copies

• Compatible with all commonly used qPCR instruments

• Includes DTI Green Master Mix for intercalator-based qPCR

• Tli RNase H Plus reduces inhibition from mRNA/cDNA hybrids in qPCR

• Eliminates the need for a separate RNase H digestion step when using low RNase H RT

DTI Green HiFid Taq HS II Premix - Protocol

DTI Green HiFid Taq HS II Premix - Certificate of Analysis

DTI Green HiFid Taq HS II Premix - Promotional Document

Results

Accurate gene expression analysis with DTI Green HiFid Taq HS II Premix

The DTI Green HiFid Taq HS II Premix enzyme efficiently and specifically amplified mouse IL-4 and the GAPDH control. Relative expression of IL-4 was higher in CRA-treated lung samples than in PBS-treated samples.

Relative expression levels of IL-4 as determined by qPCR. Fold expression levels of IL-4 (compared to GAPDH) in mouse lung samples from mice treated with either PBS or CRA.

Conclusions

Upregulation of the cytokine IL-4 is associated with allergies. It was expected that expression of IL-4 would increase during a CRA-induced model of allergic airway inflammation. Indeed, using DTI Green HiFid Taq HS II Premix, a relative increase in IL-4 gene expression was accurately detected in lung samples from mice that were sensitized and challenged with CRA. DTI Green HiFid Taq HS II Premix provided sensitive and accurate detection of gene expression.

Methods

Total RNA was purified from the lungs of C57BL/6 mice that were sensitized and challenged with either PBS (two mice) or CRA (three mice). RNA was extracted using the RNeasy Mini Kit (Qiagen), and cDNA was synthesized from 1 µl of the extracted RNA using Superscript II RT enzyme (Thermo Fisher Scientific) according to the manufacturers' protocols. Two microliters of the reverse transcription reaction (equivalent to 20 ng total RNA per 20-µl qPCR reaction) were used for qPCR with DTI Green HiFid Taq HS II Premix according to the recommended protocol. qPCR reactions were run in triplicate using probes for mouse IL-4 and GAPDH on an Applied Biosystems 7500 Real-Time PCR System. The relative gene expression level of IL-4 was determined using the delta/delta Ct method.

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