QuantiFast Pathogen RT-PCR +IC Kit (400)

Protocol tips

Upstream tips	Protocol tips	Downstream 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.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing PCR Quantitative real-time PCR - Viral using QuantiFast Pathogen RT-PCR +IC Kit (400) from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for QuantiFast Pathogen RT-PCR +IC Kit (400) below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing PCR Quantitative real-time PCR - Viral using QuantiFast Pathogen RT-PCR +IC Kit (400) from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.