What type of study is this?

This is a In Vitro Study study.

August 17, 2025

Automated high-throughput RT-PCR v2

AVAlex VeithUniversity of Wisconsin–Madison ASAnna L. ShenUniversity of Wisconsin–Madison CBChristopher BradfieldUniversity of Wisconsin–Madison

Key Points

Automated high-throughput rt-pcr yields consistent results with minimal variability across technical replicates, ensuring reliability.
The method dispenses 2 μL of diluted RNA and 8 μL of master mix for 10 μL reactions on a 384-well PCR plate, enhancing throughput.
Setup involves a pre-written template program tailored for multiple gene expression targets, streamlining the process.
Efficient arrangement of diluted RNA in a 96-well plate allows for efficient handling and optimal use of resources.

Abstract

This protocol provides an outline for planning, setting up, and running an epMotion® M5073 for high-throughput RT-PCR. This protocol leverages a set of pre-written template programs to accommodate different numbers of gene expression targets. The template programs have been designed to dispense 2 μL of diluted RNA and 8 μL of one-step master mix (including primers and probes) to create 10 μL reactions on a 384-well PCR plate. Diluted RNA is to be arranged in a 96-well PCR plate for efficiency. We also provide the steps necessary to create custom RT-PCR runs for more advanced users. These steps also detail the settings used in our predefined programs. Users can expect consistent results with minimal variability between technical replicates that are quick and reliable.

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Cite This Study

Veith et al. (Wed,) studied this question.

synapsesocial.com/papers/68a36f840a429f797333209c https://doi.org/https://doi.org/10.17504/protocols.io.bp2l6dzm1vqe/v2

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