How can I visualise digitalMLPA reactions using electrophoresis?

in Sequencing

There are two methods to visualise digitalMLPA reactions or libraries: using a capillary electrophoresis device from Applied Biosystems (ABI), or using a non-denaturing system such as the Agilent TapeStation.

Important
Visualisation of digitalMLPA reactions or libraries is only recommended for troubleshooting purposes and in case of suspected reaction failure. We do not consider a check of digitalMLPA reactions or libraries necessary for routine experiments or for data analysis, but we understand that some customers may prefer to do so. Checking digitalMLPA reactions or libraries prior to sequencing will only reveal serious issues, such as the complete absence of digitalMLPA PCR products, but does not guarantee that the quality of the reaction is sufficient for analysis. Coffalyser digitalMLPA provides more detailed information about the quality of individual reactions using a large set of control probes present in every probemix.

Capillary electrophoresis with an ABI device relies on the presence of a FAM dye on one of the primers used in the digitalMLPA reactions. This can be used to visualise relevant digitalMLPA PCR products and detect complete reaction failures. Read more about the experimental setup and the expected peak patterns. SCIEX devices cannot be used for this approach because they are not suitable for the FAM label.

It is also possible to get a broad idea of the formation of digitalMLPA PCR products in reactions or libraries using traditional NGS library assessment methods, such as the Agilent TapeStation, as long as some additional guidelines are followed. Read more about the use of a TapeStation.

Related Pages

If you have any comments or suggestions on this article, please send us your feedback.

Disclaimer

The information provided in this material is correct for the majority of our products. However, for certain applications, the instructions for use may differ. In the event of conflicting information, the relevant instructions for use take precedence.