Protocol
This section describes library preparation using the Illumina Microbial Amplicon Prep kit.
The following protocol has been tested and proven with multiple viral targets including SARS-CoV-2, Mpox (formerly monkeypox), and Dengue virus. The protocol is the recommended starting point for developing a new protocol with new targets and primer sets.
This protocol is designed for library preparation in 96-well PCR plates. If the sample number is low and better suited to library preparation in 1.5 ml microcentrifuge tubes or strip tubes, modify the protocol as necessary.
Input Types
| • | RNA only inputs: Start the protocol at the Anneal RNA step. |
| • | DNA only inputs: Start the protocol at the Amplicon PCR step. |
| • | RNA and DNA inputs: For DNA and RNA purified separately from the same starting samples, start the protocol at the Synthesize First Strand cDNA step using the RNA input. At the Amplicon PCR step, combine the cDNA and purified DNA. For optimal results, you may prefer to isolate both DNA and RNA from a sample (for example, RNA and DNA virus targets). |
The protocol provides reminders and specific changes necessary when using these input types. Some steps may require optimization, and possible optimization points are highlighted throughout the protocol.
