Ensure reproducibility and repeatability with optimized master mixes
By: Roche Life Science
Posted: | Lab Life - Real-Time PCR
Reproducibility has always been a scientific hurdle. While the basic details of an experiment are published in a manuscript, each lab can potentially interpret these instructions differently. As a result, a 2016 online survey by the journal Nature discovered that more than 70% of researchers have failed to reproduce another researcher’s results using the information given. Perhaps worse, more than half of the 1,576 respondents struggled to reproduce their own experiments.
In May 2017, Nature announced its intention to accompany all manuscript submissions with a new reporting summary document. This document goes above and beyond the previous checklist, which was designed to circumvent the problems that often lead to irreproducibility.
The new reporting summary document will require details about the experiment’s design, tools and reagents used, and other methodological details that would improve the likelihood of being able to reproduce and repeat the findings from lab to lab. The journal editors believe that this will encourage transparency, and help reviewers, editors, and readers to assess the quality of the experiments, and the resulting analysis.
How can your lab ensure reproducibility?
One easy way to improve the reproducibility and repeatability of results is the use of optimized master mixes, mixes that minimize the variables that occur among researchers and laboratories. These master mixes use a precisely calibrated blend of reagents to minimize the variability between test runs, either in-house or externally.
These optimized master mixes offer several benefits for your lab:
- They’re ready to use: Master mixes are a ready-to-use, 2x concentrated solution
- Contains all reagents: Each master mix contains DNA Polymerase, dNTPs, and buffers, each in optimum concentrations
- Quick setup: When using a master mix, the only variables are template and primers
- Minimize contamination risk: Fewer pipetting steps reduces the risk of contamination in your experiment
- Specificity and sensitivity: Consistent formulations deliver the sensitivity and specificity needed to ensure accuracy, time after time
- Consistent results: Master mixes are specifically optimized to deliver consistent, reproducible, and repeatable results
The right master mix increases throughput, accuracy
Choosing the right master mix for your device and application can increase your throughput, enabling you to run more samples with less preparation time. Test results will be more accurate, with less risk of contamination, and improved consistency from experiment to experiment. In addition to aiding reproducibility, this also improves the overall efficiency of the lab, making optimized master mixes a cost-effective investment.
How to choose the right master mix
When selecting a master mix, consider the following:
- Sample type: DNA or RNA
- Target sequence: The selected sequence for amplification
- Application: End-point PCR, qPCR, or RT-PCR
- Device: While generic, cross-platform master mixes are available, it’s often best to use a mix that is optimized for the specifics of your device
Selecting an optimized master mix will improve solution consistency and minimize contamination to improve accuracy in laboratory testing. This ensures consistent results to improve the reproducibility and repeatability of your results in published research.