QuantiTect Probe RT-PCR Kit (1000)

PCR Quantitative real-time PCR - Viral

Experiment
PCR Quantitative real-time PCR - Viral
Product
QuantiTect Probe RT-PCR Kit (1000) from Qiagen
Manufacturer
Qiagen

Protocol tips

Protocol tips
Follow manufacturer's instructions

Publication protocol

TaqMan RT-rtPCR assays: RT-rtPCR protocol: RT-rtPCR was performed using a total volume of 25 µL on 2 µL of each RNA dilution and 23 µL of the master mix components from each tested kit. The concentrations of the forward and reverse primers and the Lys-Gt1 probe were optimized for each tested kit.21
All commercial kits were optimized with regard to the reverse transcription duration according to the manufacturer’s instructions (Table 1). The same PCR thermal cycling conditions (Table 1) were applied for the 6 kits tested (i.e., 45 cycles of 30 s at 94°C, 30 s at 55°C, and 30 s at 72°C). Real-time PCRs were performed on 2 real-time thermocyclers: Rotor-Gene Q MDx (RG, Qiagen) and Mx3005P (Agilent Technologies, Les Ulis, France). PCR runs were performed in independent assays on multiple days by one trained laboratory technician. Preparation of PCR master mixes was performed by manual pipetting.

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Discussion

Discussion

5 years ago

Author: Germany

What is the optimal concentration for primers in qPCR?

What is the optimal concentration for primers in qPCR? My total volume is 20μl per reaction.

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Papers

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Manufacturer protocol

Download the product protocol from Qiagen for QuantiTect Probe RT-PCR Kit (1000) below.

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