PCR Quantitative real-time PCR - Bacterial DNA

A PCR reaction consists of the template DNA, two primers covering the amplification site, an enzyme, and buffers. A quantitative, real-time PCR reaction typically includes all of that plus a probe that can be detected fluorescently as the reaction runs, with no gel required. for detection. However, non-specific product amplification and primer-dimer formation during set-up are major causes of PCR failure. Nevertheless, high-quality DNA polymerase and optimize reaction buffers will certainly lead to a successful PCR reaction.

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5 years ago

5 years ago by Jana Fassbinder 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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Found 6 matching solutions for this experiment

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
PCR plates, tubes and pipette tips should be UV sterilized for 20-30 mins
Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
PCR plates, tubes and pipette tips should be UV sterilized for 20-30 mins
Fast SYBR™ Green Master Mix

Thermo Fisher Scientific

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
CR plates, tubes and pipette tips should be UV sterilized for 20-30 mins
SYBR™ Green PCR Master Mix

Thermo Fisher Scientific

Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp
Protocol tips
PCR plates, tubes and pipette tips should be UV sterilized for 20-30 mins
Upstream tips
Amplicon size should be consistent for each target sequence and limited to approximately 65 - 100 bp.
Protocol tips
PCR plates, tubes and tips should be UV sterilized for 20-30 mins
Protocol tips
Test different primer/probe concentrations and extension temperatures in order to achieve maximum replication efficiency.
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