One-step qPCR Master Mix
Code No. QRT-101 100 reactions/Code No. QRT-201 100 reactions
This product is a 2 × Master Mix for "1-step real-time PCR" using a thermostable DNA polymerase derived from Thermus thermophilus (Tth) HB8 (1). Tth DNA polymerase exhibits reverse transcriptase activity in the presence of Mn2+ ions. This system allows for "1-step real-time PCR", including reverse transcription and PCR steps. RNA-direct™ Realtime PCR Master Mix is applicable for TaqMan™ assay or hybridization probe assay, in combination with each probe. RNA-direct™ SYBR Green™ Realtime PCR Master Mixcan be applied to an intercalation assay with SYBR® Green I.
<RNA-direct Realtime PCR Master Mix>
TaqMan™ assays or hybridization probe assays
<RNA-direct SYBR® Green Realtime PCR Master Mix>
Intercalation assay with SYBR® Green I
Store at -20ºC
This reagent includes the following components for 100 reactions, with a total of 50 µL per reaction. All reagents should be stored at -20ºC.
<QRT-101>
RNA-direct™ Real-time PCR Master Mix | 0.5 mL x 5 |
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50 mM Mg(OAc)2 | 0.5 mL x 1 |
<QRT-101>
RNA-direct™ SYBR® Green Realtime PCR Master Mix | 0.5mL x 5 |
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50mM Mg(OAc)2 | 0.5mL x 1 |
Correlation between the 1-step method (RNA-direct™ SYBR® Green Realtime PCR Master Mix [Code No. QRT-201]) and conventional 2-step method (ReverTra Ace™ [Code No. TRT-101] (highly efficient reverse transcriptase)->SYBR Green™ Realtime PCR Master Mix [Code No.QPK-201]) was evaluated based on the expression of Protein kinase δ (PKCδ) mRNA. The results from the 1-step method were found to be highly correlated with those of the 2-step method (Fig. 1).
The Y-axis indicates the relative quantitation value of Protein kinase δ (PKCδ) mRNA
Yellow bar (left): 1-step method using 50 ng total RNA
Yellow bar (right): 1-step method using 200 ng total RNA
Blue bar: 2-step method
Amplification of G3PDH mRNA was detected using serially diluted poly(A)+ RNA (10n dilution) with SYBR® Green Realtime PCR kits including Tth DNA polymerase. The RNA-direct™ SYBR® Green Realtime PCR Master Mix [Code No. QRT-201)] showed greater sensitivity and signal intensity than the other kit (company A).