QuantiFast Pathogen RT-PCR +IC Kit (400)

PCR Quantitative real-time PCR - Viral

Experiment
PCR Quantitative real-time PCR - Viral
Product
QuantiFast Pathogen RT-PCR +IC Kit (400) from Qiagen
Manufacturer
Qiagen

Protocol tips

Publication protocol

The one-step real-time RT-PCR was developed using the QuantiFast pathogen RT-PCR +IC kit (Qiagen, Hilden, Germany) and was carried out in an ABI 7300 real-time PCR system (Applied Biosystems, Foster City, CA, USA). The real-time RT-PCR conditions were as follows: 5 μl of extracted RNA, 5 μl of 5× QuantiFast pathogen RT-PCR master mix, 0.25 μl of QuantiFast pathogen RT mix, 100 nM primers and probe (0.03 μl of stock 80 μM), 2.5 μl of 10× internal control RNA, 2.5 μl of 10× internal control assay, 0.50 μl of High-ROX dye solution, and RNase-free water to adjust the volume to 25 μl. The cycling conditions for the amplification were as follows: reverse transcription at 50°C for 20 min, an initial PCR activation step at 95°C for 5 min, and 45 cycles of two-step cycling for 10 s at 95°C and for 30 s at 60°C.

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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 QuantiFast Pathogen RT-PCR +IC Kit (400) below.

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