We’ve put together a new small RNA (microRNA) sequencing guide describing considerations all new users should make before undertaking a small RNA sequencing project. One of the first considerations is determining the number of reads you need. This usually depends on whether you’re interested in differential small RNA expression or if you’re trying to discover new microRNAs. Once you know the number of reads you need per sample, consider the following factors before and after library preparation:
- Should you start with total RNA or isolated small RNA?
- How much material should you start with?
- What’s the minimum quality of total RNA acceptable for microRNA library preparation and sequencing?
- How will small RNA ligation bias affect my results?
- How can I minimize adapter dimers to improve read mapping and general usability of my sequencing reads?
- How many samples can I multiplex or pool together in a single sequencing lane?
- What sequencing read length should I choose for microRNA or small RNA sequencing studies?
The guide also includes recommendations for getting accurate per sample pricing and turnaround times.
Small RNAs play a big role in regulating the translation of target RNAs through RNA to RNA interactions and have been shown to offer potential as biomarkers in diagnostic applications. Sequencing promises to be a useful tool in unraveling the roles of these short non-coding RNAs. We look forward to working with you on your next microRNA project.