qPCR and ddPCR

Key Points

qPCR:

ddPCR:

Review Questions

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Overview

Quantitative polymerase chain reaction (qPCR) is used to measure DNA replication in real time. In the lab, qPCR is generally used to measure RNA expression. You convert RNA to cDNA with reverse transcriptase, and use target-specific probes (just primers that fluoresce) to amplify a transcript of interest. By tracking how quickly the cDNA of interest amplifies, you can infer how much of that RNA was initially present in the sample Digital droplet polymerase chain reaction (ddPCR) works just like qPCR but, first, the sample to be measured is separated into thousands of droplets that can be amplified and analyzed separately. This allows for many replicates of the same sample, which increases statistical power and can help to detect RNA transcripts at lower expression levels.

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Additional Considerations

Articles

Droplet Digital PCR (ddPCR) TechnologyBio-Rad Laboratories

https://www.nature.com/articles/s41598-017-02217-x.pdfwww.nature.com

A paper describing differences between qPCR and ddPCR

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Question Answers